蛋白纯化策略 - Others

重组蛋白的纯化

AMERSHAM-BIOSCIENCES    重组蛋白手册The Recombinant Protein Handbook   (pdf-I)   (pdf-II)
PIERCE:   融合蛋白纯化手册Fusion Protein Products   (pdf)
SIGMA:   蛋白纯化和检测选择指南 Detection and Purification Selection Guide   (pdf-I)    (pdf-II)

His-Tagged
融合蛋白的纯化

       NOVAGEN:   pET purification manual (pdf)
        NOVAGEN: Ni-NTA Hi-Bind Resins Protocols   (pdf)
        NOVAGEN: His-Tag GST-Tag purification and Detection tools   (pdf)
        QIAGEN : Ni-NTA purification manual (pdf)
        QIAGEN : NiNTA cell lysis under nature conditions (pdf)
        MERCK-NOVAGEN:  Metal Chelate Affinity Chromatography. Ni-MACTM, Co-MACTM and u-MACTM Metal Affinity Chromatography (MAC) Resins
                 and Cartridges   (pdf)   (pdf-II)    (pdf-III)
        IBA  Tools for protein expression & purification. 6xHis-tag & Ni-NTA technology: The optimal partner for Strep-tag in double tag proteins   (pdf)
        IBA: Expression and purification of proteins using Strep-tag and/or 6xHistidine-tag. A comprehensive manual.    (pdf)
        IBA  Mammalian expression and purification system using Strep-tag and/or 6xHistidine-tag   (pdf)
        CLONTECH: Talon Resin Protocol (pdf)
        CLONTECH: Talon Products (pdf)
        CLONTECH:  BD TalonTM Metal Affinity Resins User Manual   (pdf)
        CLONTECH:   Purification in the presence of Beta Mercaptoethanol   (pdf)
        CLONTECH:   Protein Purification Products   (pdf)
        CLONTECH:  BD HAT Protein Expression and Purification System User Manual   (pdf)
        AMERSHAM-BIOSCIENCES Hi Trap Chelating (pdf)
        AMERSHAM-BIOSCIENCES Hi Trap Chelating Application Note   (pdf)
        AMERSHAM-BIOSCIENCES   Chelating Sepharose Fast Flow Instructions   (pdf)
        AMERSHAM-BIOSCIENCES   His GraviTrap prepacked, single-use column for purification of histidine-tagged proteins by immobilized metal
                affinity chromatography (IMAC) without any need for a purification system. Negligible nickel leakage and is compatible with denaturing and
                reducing agents as well as a wide range of additives. Short purification times. High protein binding capacity, ≈ 40 mg/column. Purifies
                unclarified samples   (pdf)
        AMERSHAM-BIOSCIENCES    His SpinTrapTM is a prepacked, single-use spin column for purifying histidine-tagged proteins by immobilized
                metal ion affinity chromatography (IMAC). The resin has high protein binding capacity, low nickel ion (Ni2+) leakage, and excellent compatibility
                with denaturing agents plus a wide range of additives. His SpinTrap is used with a standard microcentrifuge and one purification run takes
                approx. 10 min. His SpinTrap columns allow direct purification of unclarified cell lysates.   (pdf-I)   (pdf-II)
        AMERSHAM-BIOSCIENCES   HisTrapTM FF crude. This pre-prepacked column is intended for preparative puri?cation of
                histidine-tagged recombinant proteins from unclari?ed lysate without precentrifugation and ?ltration of the sample. Leakage of Ni2+
                is very low. The medium is compatible with all commonly used aqueous buffers, reducing agents, denaturants such as 6 M Gua-HCl
                and 8 M urea, and a range of other additives (pdf)
         AMERSHAM-BIOSCIENCES    Ni Sepharose and IMAC Sepharose selection guide   (pdf)
         AMERSHAM-BIOSCIENCES     IMAC Sepharose High Performance. Possible to charge with various metal ions for optimized selectivity.
                High protein binding capacity. Compatible with commonly used reducing agents, such as DTT, DTE, TCEP, and β-mercaptoethanol.
                Stable in a wide range of denaturants, detergents, and buffer systems. (pdf)
        AMERSHAM-BIOSCIENCES     Ni Sepharose High Performance   (pdf)   (pdf-II)   
        AMERSHAM-BIOSCIENCES    Article: Addition of imidazole during binding improves purity of histidine-tagged proteins   (pdf)

         ACTIVE MOTIVE: Ni-Ted Manual   (pdf)
        AFFILAND: Metal Purification
        VIVASCIENCE:  Vivapure Metal Chelate Mini Spin Columns   (pdf-I)   (pdf-II)   (pdf-III)   (pdf-IV)   (pdf-V)   (pdf-VI)    (pdf-VII)    (pdf-VIII)
         Small scale His-Tag fusion protein purification under nature conditions
        Sm all scale His-Tag fusion protein purification under denaturative conditions
DYNAL  Dynabeads TalonTM   (pdf)        Ni-NTA: Handling and Reuse        SIGMA:  His Selected Nickel Affinity Gel   (pdf)
        PROMEGA:  MagneHisTM Protein Purification System   (pdf)      
        HISPANAGAR  Chelate Agarose Beads   (pdf-I) (pdf-II)   (pdf-III)   
        SARTORIUS   Sartobind® IDA 75 - A Separation Technology Based on Metal Chelate Membrane Adsorbers - Operating Instructions   (pdf)

Maltose麦芽糖结合蛋白
NEW ENGLAND BIOLAB: pMAL protein Fusion and Purification System (pdf-I)    (pdf-II)
         Small scale MBP-fusion protein purification

       Crystal Structures of Fusion  Proteins with Large-Affinity tags.  D.R.Smyth et.al.  Protein Science (2003), 12: 1313-1322   (pdf)

Calmodulin
钙调蛋白结合蛋白
     STRATAGENE: CBP Calmodulin-Binding Peptide Affinity Tag System   (pdf-I)   (pdf-II)   (pdf-III)

Intein
内蛋白介导纯化
     NEW ENGLAND BIOLAB: INTEIN-TM System   (pdf)
      NEW ENGLAND BIOLAB: INTEIN-TWIN System   (pdf)
T7.Tag
     NOVAGEN T   7.Tag Affinity Purification kit   (pdf)
Cellulose
纤维素结合位点
NOVAGEN  CBIND Kits (pdf)
NusA Protein
NOVAGEN   Expression of Soluble Heterologous Proteins via Fusion with NusA Protein - Article (pdf)

Biotinylated
生物素酰化融合蛋白

        PROMEGA    PinPointTM Xa Protein Purification System  The System is designed for the production and purification of fusion proteins that
                are biotinylated in vivo. Biotinylated fusion proteins are produced in E. coli and are affinity-purified using the SoftLinkTM Soft Release
                Avidin Resin. This proprietary resin allows elution of the fusion protein under nondenaturing conditions. The PinPointTM Vectors
                feature the encoded endoproteinase Factor Xa  proteolytic site that provides a way to separate the purification tag from the native protein.  (pdf)   

FLAG

        SIGMA:  Detection and Purification - FLAG®  蛋白纯化检测验证系统  (pdf)

Strep-Tag   

        IBA:  Expression and purification of proteins using Strep-tag and/or 6xHistidine-tag. A comprehensive manual. The Strep-tag II is a short peptide
                 (8 amino acids, WSHPQFEK), which binds with high selectivity to Strep-Tactin, an engineered streptavidin. Elution of purified recombinant
                  protein is performed by desthiobiotin.   (pdf)
        IBA  Tools for protein expression & purification. Strep-tag® technology and 6xHis-tag & Ni-NTA technology: Double tag protein expression
                 and purification   (pdf)
        IBA  Mammalian expression and purification system using Strep-tag and/or 6xHistidine-tag   (pdf)
        IBA  One-STrEP Kit. One-STrEP-tag Purification for the Isolation and Identification of Protein Complexes in Mammalian Cells.
                 A comprehensive manual   (pdf)
       NOVAGEN  Strep

·
Tactin® Purification Kits. StrepTactin protein is a streptavidin derivative developed for optimal StrepTag II binding. The

                 binding affinity of Strep

·
Tag II for StrepTactin is approximately 100 times higher than for streptavidin. The purified target protein is competitively

                 eluted with 2.5 mM desthiobiotin, an analog of biotin that reversibly binds Strep

·
Tactin. (pdf-I) (pdf-II)

       QIAGEN:   Two-Step Affinity Purification System Handbook - For expressing, purifying, and detecting proteins carrying a 6xHis
                 and Strep-tag II. After elution from the Ni-NTA matrix using imidazole, recombinant proteins (which also carry the Strep-tag II
                 epitope) are loaded directly onto a Strep-Tactin matrix and eluted using either biotin or desthiobiotin.    (pdf)
        STRATAGENE:    VariFlexTM bacterial protein expression system. Include three different solubility enhancement tags (SETs) which are
                 designed to increase protein solubility, the streptavidin binding peptide (SBP) purification tag, and a tag that allows for rapid soluble protein
                 quantification (Q-tag)    (pdf)

Fluorescent Protein荧光蛋白
         BD BIOSCIENCES   BD Living ColorsTM AcGFP1 Fluorescent Protein. A novel monomeric green fluorescent protein for fusion tag
                    applications. Ideal for multicolor applications in flow cytometry and fluorescence microscopy.   (pdf)   
        [url=file:///C:/My%20Documents/site/www.bdbiosciences.com]BD BIOSCIENCES[/url]   BD Living ColorsTM DsRed-Monomer Fluorescent Protein. A novel monomeric red fluorescent protein for fusion tag
                    applications. Ideal for multicolor applications in flow cytometry and fluorescence microscopy. (pdf)

Halo tag fusion

       PROMEGA:   The HaloTagTM Interchangeable Labeling Technology is a novel tool for imaging live or fixed mammalian cells that express the
                HaloTagTM Protein or protein fusions, analyzing post-translational modification of labeled fusion proteins, and isolating proteins and protein
                complexes. The technology is based on efficient formation of a covalent bond between a specially designed reporter protein encoded by the
                HaloTagTM pHT2 Vector and a specific ligand in living cells, in solution or on a solid support.  The HaloTagTM Ligand can carry a variety of
                functionalities, including fluorescent labels, affinity handles and attachments to a solid phase. The covalent bond forms rapidly under general
                physiological conditions, is highly specific and essentially irreversible, yielding a complex that is stable even under stringent conditions. The open
                architecture of the technology enables use of different ligands. Technical Manual. (pdf)
       PROMEGA:   HaloLinkTM Resin. The HaloTag technology comprises the HaloTag polypeptide, which can be fused to a protein of interest using
                the HaloTag Vectors, and a system of interchangeable synthetic ligands that covalently bind to the HaloTag polypeptide. HaloLink Resin, allows
                covalent and oriented surface immobilization of HaloTag fusion proteins. The HaloLink Resin can be used in a variety of applications including
                enzyme immobilization, detection of protein:protein interactions and purification of fusion proteins using protease cleavage. (pdf)

Solubility enhancement tags (SETs)

        STRATAGENE:    VariFlexTM bacterial protein expression system. Include three different solubility enhancement tags (SETs) which are
                  designed to increase protein solubility, the streptavidin binding peptide (SBP) purification tag, and a tag that allows for rapid soluble protein
                  quantification (Q-tag)    (pdf)

  SUMO

        INVITROGEN:   The ChampionTM pET SUMO Protein Expression System utilizes a small ubiquitinlike modifier (SUMO) to allow expression,
                 purification, and generation of native proteins in E. coli. SUMO fusions may increase the expression of recombinant proteins and enhance the
                 solubilityof partially insoluble proteins. In addition, the tertiary structure of the SUMO protein is specifically recognized and cleaved by a
                 ubiquitin-like protein-processing enzyme, SUMO Protease resulting in the production of native protein.   (pdf)
        LIFESENSORS  SUMO Gene Fusion Technology - New Methods for Enhancing Protein Expression and Purification in Eukaryotes   (pdf)  
        LIFESENSORS  SUMO Gene Fusion Technology - New Methods for Enhancing Protein Expression and Purification in Prokaryotes   (pdf)   
        LIFESENSORS  SUMO Gene Fusion Technology - New Methods for Rapid Recombinant Peptide Expression and Purification   (pdf)

Cold Expression

          TAKARA  pCold TF DNA Vector is a fusion cold shock expression vector that expresses Trigger Factor (TF) chaperone as a soluble tag
                    (48 kDa) which facilitates co-translational folding of newly expressed polypeptides. The vector has a His-Tag sequence, and a multicloning site
                    (MCS). A lac operator is inserted downstream of  the cspA promoter to ensure strict regulation of expression. Additionally, recognition sites
                    for HRV 3C Protease, Thrombin, and Factor Xa are located between TF-Tag and the Multiple Cloning Site (MCS) and function to facilitate
                    tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts. The pCold TF DNA Vector provides cold
                    shock technology for high yield protein expression combined with Trigger Factor (chaperone) expression to facilitate correct protein folding,
                    thus enabling efficient soluble protein production for otherwise intractable target proteins.   (pdf-I)   Chaperone Plasmid Set / Chaperone
                    Competent Cells (pdf-II)   

Cleavage of Recombinant Proteins融合蛋白酶切裂解

Factor Xa

Factor Xa Cleavage of a MBP fusion protein
       NOVAGEN: Factor-Xa kit   (pdf)
       QIAGEN:   Factor Xa  and Factor Xa Removal resin   (pdf)
Thrombin
Thrombin Cleavage of a GST fusion protein
       NOVAGEN:  Thrombin kit    (pdf)
Enterokinase
     NEW ENGLAND BIOLABS:   Enterokinase (pdf)
      NOVAGEN:  Enterokinase cleavage capture kit   (pdf)
     ROCHE:    Enterokinase   (pdf)
PreScission
     AMERSHAM-BIOSCIENCES Application Note   (pdf)
      AMERSHAM-BIOSCIENCES Life Science News   (pdf)

   TAGZymeQIAGEN:   His-tag Removal by exoproteolytic Digestion   (pdf)

   TEV-Protease
INVITROGEN- LIFE TECHNOLOGIES  TEV-Protease (pdf)
      Mohanty A. et.al. (2003) Inhibition of tobacco etch virus protease activity by detergents. Protein Expression and Purification 27: 109-114   (pdf)
       CARDIFF UNIVERSITY - MOLECULAR CELL BIOLOGY - EHRMAN LAB   TEV NIa protease
HRV 3C Protease

        NOVAGEN    HRV 3C Protease   (pdf)      

SUMO   

        INVITROGEN   SUMO Protease   (pdf)   
        LIFESENSORS  SUMO Protease 1   (pdf)    (pdf-II)

Affinity Chromatography亲和层析

Activated Resins偶联用凝胶
   AMERSHAM-BIOSCIENCES Affinity Manual  (pdf)
      AMERSHAM-BIOSCIENCES Affinity Columns and Media  (pdf)  
            HiTrap NHS-activated HP, NHS-activated Sepharose 4 Fast Flow (10-atom spacer arms) is designed for the covalent coupling
            through the primary amine of a ligand and is the first choice for the preparation of immunospecific media.
            CNBr-activated Sepharose offers a well-established option for the attachment of larger ligands and as an alternative to NHS-activated
            Sepharose. Proteins, peptides, amino acids or nucleic acids can be coupled to CNBr-activated Sepharose, under mild conditions, via
            primary amino groups or similar nucleophilic groups.
            EAH Sepharose  4B for coupling small ligands containing free acarboxyl groups via a 10-atom spacer arm. Carbodiimide as the coupling
            reagent. Phenolic groups may be attached through others couple reagents. Thiol derivatives can couple carboxyl groups in the presence of
            carbodiimide and the thiol ester bond may be cleaved specifically using hydroxylamine, thus providing a simple and gentle method for
            eluting the intact ligand-protein complex.
            ECH Sepharose 4B for coupling small ligands containing free amino groups via a 9-atom spacer arm. Carbodiimide as the coupling reagent.
            Epoxy-activated Sepharose 6B for coupling through hydroxy, amino or thiol groups via a 12-carbon spacer arm. It is particularly useful for
            coupling small ligands such as choline, ethanolamine and sugars. The pre-activated matrix is formed by reacting Sepharose 6B with the
            bis oxirane, 1,4 bis-(2,3-epoxypropoxy-)butane. Free oxirane groups couple via stable ether bonds with hydroxyl-containing molecules
            such as sugars, via alkylamine linkages with ligands containing amino groups, and via thioether linkages with ligands containing thiol groups.
            Thiopropyl Sepharose 6B for coupling through a thiol group many types of small ligands
                  

·
Heavy metal ions and derivatives can be used as ligands to react with thiol groups forming mercaptides.

·
Alkyl or aryl halide ligands give thioether derivatives.

·
Ligands containing C=O, N=N and, under certain conditions, C=C bonds undergo addition reactions.

            The hydroxypropyl group acts as a small spacer arm. Ligands containing amino groups can be attached to Thiopropyl Sepharose 6B
            or Activated Thiol-Sepharose 4B by multi-point attachment or coupling through a small number of groups using the heterobifunctional
            thiolating reagent, SPDP. The coupled molecules may be recovered by eluting with a reducing agent.
      AMERSHAM-BIOSCIENCES CNBr Activated Sepharose Data File  - React with primary amines present on proteins, antibodies
            and other molecules.  (pdf)
      AMERSHAM-BIOSCIENCES   NHS Activated Sepharose Data File -NHS (N-hydroxysuccinimide) coupling forms a chemically
            stable amide bond with ligands containing primary amino groups. NHS-activated Sepharose® 4 Fast Flow provides a spacer arm
            and is therefore particularly suitable for immobilising small protein and peptide ligands.  (pdf)   
       AMERSHAM-BIOSCIENCES   Recommended carbodiimide coupling procedure for CH Sepharose® 4B and AH Sepharose® 4B (pdf)
      BIO RAD   Affi-Gel 10 and Affi-Gel 15 activated immunoaffinity supports (N-hydroxysuccinimide esters of a derivatized crosslinked agarose),
            with a 10 and 15-atom spacer arm respectively, that offer rapid, high efficiency coupling for all ligands with a primary amino group aqueous
            or non-aqueous solution.     (pdf)
      HISPANAGAR  Amino Ethyl and Glyoxal Agarose Beads  (pdf-I)   (pdf-II)   (pdf-III)   
            Glyoxal Agarose Beads  includes a broad selection of resins for the immobilization of biomolecules through amino groups (enzymes,
            antibodies, etc). There are resins, with either 4% or 6% agarose concentration, and with various degrees of activation (very high/ high/low).
            This permits the immobilization of biomolecules in different size ranges.
            Aminoethyl Agarose Beads covalently bind the agarose to the acid groups of the amino acid of the target biomolecules. There are resins
            with agarose concentration of 4% or 6% and with a range of degrees of activation.
      JENA BIOSCIENCE   Immobilized Nucleotides for Affinity Chromatography    (pdf)   Immobilized nucleotides provide a convenient
            and rapid one-step purification procedure for a large number of proteins such as kinases, GTPases, chaperones, motor proteins, and others.
            Jena Bioscience provides NTPs and dNTPs that are linked to the matrix at various positions of sugar, base or phosphate moiety of the
            nucleotide; different types and lengths of linkers and different types of chromatography material ranging from bulk material to pre-made
            columns that fit any machine
      MERCK  Fractogel EMD  Amino   (pdf)   For immobilization of ligands with carboxy-functionional groups.
      MERCK  Fractogel EMD  Epoxy (M)   (pdf)   Suitable for coupling of low molecular weight ligands and pH stable proteins. The epoxyde reacts
            with primary amino groups, hydroxyl, and sulfhydryl groups. The resulting affinity matrix is very stable, due to the ether bonding of the ligand.      NOVAGEN   Preactivated Resins  (pdf)
           PreACTTM Agarose ALD is an activated chromatography matrix for simple and efficient immobilization of small ligands to large
            proteins under mild physiological conditions. Is supplied in an activated form containing bound reactive aldehyde groups. During
            ligand coupling, aldehydes react with primary amines on the ligand to form a Schiff-base. In a further reaction, the Schiff-base linkage
            is selectively converted into a stable, secondary amine linkage.
            PreACT Fractogel AZL is an activated resin with a high density of tentacle-bonded azlactone reactive groups suitable for the
            coupling of proteins under physiological conditions.  The azlactone ring is opened by nucleophilic attack of appropriate groups
            present on the protein surface. Upon reaction with amines, a stable amide bond forms between the former carbonyl function of the azlactone.
            PreACT Fractogel EPX is an activated resin for the immobilization of low molecular weight amine bearing ligands or alkaline-stable proteins.
            Bonded epoxide groups react with primary amino, hydroxyl and sulfhydryl groups to form stable ether linkages.
      PIERCE:   亲和层析手册 (pdf)      PIERCE   偶联不同功能基团: Tosyl, Tresyl and Epoxy Activated Agarose. Alkylamine Beads
            Tosyl Activated Agarose Hydroxyls on the surface of cross-linked beaded agarose are reacted with p-toluenesulfonyl chloride (tosyl chloride)
            to yield a sulfonated support. These sulfonates can couple to nucleophiles, such as primary amines or thiols, to yield a stable affinity support.
            The tosylated support also couples to imidazole, or tyrosine hydroxyl groups. Tosyl Activated Agarose couples more rapidly and more efficiently
            to sulfhydryl groups than to primary amines. Sulfhydryl coupling supports are convenient when accessible free sulfhydryls exist in the protein, or
            when one can be easily generated through reduction of a disulfide bond.
            Tresyl Activated Agarose Hydroxyls on the surface of agarose are reacted with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride) to yield
            a sulfonated support.    This sulfonated support is approximately 100-fold more reactive than a Tosyl agarose support. Tresyl Activated Agarose
            can couple to nucleophiles, such as primary amines or thiols, to yield a stable affinity matrix.. The tresylated support also couples to imidazole
            and tyrosine hydroxyl groups. Tresyl Activated Agarose couples more rapidly and more efficiently to sulfhydryl groups than to primary amines.
            ImmunoPure® Epoxy-Activated Agarose Epoxide chemistry is a useful way to immobilize ligands  like proteins, carbohydrates, peptides and
            amino acids, containing nucleophiles, such as amino,thiol and hydroxyl (including phenolic) functional groups. Epoxide-activated supports are
            produced by the immobilization of bifunctional oxiranes such as 1,4-butanediol diglycidyl ether onto agarose supports, introducing a hydrophilic
            spacer arm. These activated supports have limited stability in aqueous media, so it is necessary to use them quickly after they are generated or
            rehydrated.   (pdf)
      PIERCE   CarboLink® Coupling Gel allows immobilization of glycoproteins through their oxidized carbohydrate moieties. Because
            carbohydrates are located on the Fc portion of antibody molecules, CarboLink® Gel has the advantage of orienting the antibody
            binding sites to remain unobstructed, resulting in greater purification capability.  (pdf)
      PIERCE   Coupling Proteins trough Sulfhydril groups  Pierce SulfoLink® Coupling Gel is designed to efficiently react with thiol-containing
            molecules and immobilize them through a thioether linkage. The support contains an iodoacetyl group at the end of a long spacer arm,
            which reacts with sulfhydryls through displacement of the iodine.  (pdf)
      PIERCE:    The AminoLink® Coupling Gel support is 4% cross-linked beaded agarose that has been activated to form aldehyde
            functional groups. The aldehydes react spontaneously with primary amines found in lysine residues and at the amino terminus of a
            peptide chain. Reductive amination of the resulting Schiff base then forms a stable, secondary amine linkage with minimal leakage
            of the ligand.  (pdf)
      PIERCE:   Batch and Spin Cup Methods for Affinity Purification of Proteins   (pdf)   
      PIERCE:   MicroLinkTM Protein Coupling Kit allows immobilization of small amounts (25-100 µg) of purified antibody and other
            proteins directly onto beaded agarose gel to create a permanent affinity support. The AminoLink® Plus Coupling Gel included
            in this kit contains aldehyde functional groups that react with primary amines present on antibodies and other molecules.
            Reductive amination of the resulting Schiff bases forms a stable secondary amine linkage with minimal leakage. (pdf)
      PIERCE:   MicroLinkTM Peptide Coupling Kit. For sulfhydryl-containing peptide or protein. This kit uses UltraLink® Iodoacetyl Gel
            that reacts specifically with free sulfhydryls to form a stable thioether linkage. The support contains a 15-atom spacer that reduces
            steric hindrance, making binding interactions with the coupled molecule efficient. (pdf)
      PIERCE:   Ultra Link Immobilized Carboxy for Inmobilization of Peptides using EDC. Useful for coupling primary amine containing ligands
            like small peptides   (pdf)
      PIERCE:   UltraLink® Iodoacetyl Gel binds specifically to sulfhydryls when used under specific conditions with the iodoacetyl alkylating agent.
            The 15-atom spacer arm is especially ideal for conjugating small peptides to the support. The support will immobilize numerous types of
            molecules with a free sulfhydryl including antibodies, other proteins and peptides.   (pdf)      PIERCE:   Ultra Link Biosupport Medium with Azlactone Groups. Couples nucleophiles on ligands via a ring opening reaction to
            attach the ligand to the support through stable covalent linkages. Amino-functional ligands will form stable amide bonds at the end
            of a five-atom spacer.   (pdf)
      SARTORIUS  Sartobind® Epoxy 75 - A Microporous Coupling Membrane for Affinity Chromatography - Operating Instructions -
            Any molecule containing amino-, hydroxyl- or thiol-groups may be immobilized by covalent coupling to the epoxy-activated membrane.
            The membrane is fitted into a filter holder with Luer Lock connectors for easy handling to quickly couple biomolecules like proteins
            or peptides covalently.  (pdf)
      TOSOH BIOSCIENCE   Affinity Chromatography Activated Resins   (pdf)
            TOYOPEARL AF-Tresyl-650M:  For coupling of proteins through amine and thiol groups at slightly alkaline pH (7.0-8.0)
            TOYOPEARL AF-Epoxy-650M: Useful for attaching low MW ligands at high densities and for immobilization of carbohydrates or glycoproteins
            TOYOPEARL AF-Formyl-650M: (aldehide bearing) For coupling of proteins through amine groups at slightly alkaline pH (7.0-9.0)              TOYOPEARL AF-Amino-650M:  For coupling ligands through their carboxylate groups (peptide bond formation) or aldehyde groups
            (reductive amination) present in carbohydrates and glycoprotein ligands, or introduced into the ligand by mild periodate oxidation. Reaction at
            slightly acidic pH (4.5-6.0)              TOYOPEARL AF-Amino-650M: Carbodiimide mediated coupling reaction to amino groups of proteins or low MW ligands. Reaction at
            slightly acidic pH (4.5-6.0)  
      VIVASCIENCE:  Vivapure Epoxy Coupling Kit - Mini Spin Columns - Vivapure Epoxy Mini spin columns provide a membrane to
            which any desired protein, containing amino-, sulfhydryl-, or hydroxyl-groups on its surface is coupled covalently.  (pdf)
     

       Antibody Purification抗体纯化
      

        AMERSHAM-BIOSCIENCES   抗体纯化手册Antibody  Purification Manual   (pdf)   
         AMERSHAM-BIOSCIENCES   Application Note: Rapid optimisation and development of an automated two-step purification procedure for
                           monoclonal IgG antibodies   (pdf)   
        AMERSHAM BIOSCIENCES   Capto adhere: a strong anion exchanger with multimodal functionality ionic interaction, hydrogen bonding and
             hydrophobic interaction. Capto adhere is designed for post Protein A purification of monoclonal antibodies. Removal of leached Protein A,
            aggregates, host cell proteins, nucleic acids and viruses from monoclonal antibodies is performed in flowthrough mode at where the antibodies
            pass directly through the column while the contaminants are adsorbed.   (pdf-I)   (pdf-II)   (pdf-III)
        BIORAD    Affi-Prep® Protein A Matrix Instruction Manual   (pdf)
        BIORAD    Affi-Gel® Protein A MAPS® II Kit - Instruction Manual  [url=] (pdf)
[/url]        BIORAD    Econo-Pac® Protein A Kit Protein A Columns - Instruction Manual   (pdf)
        BIO RAD   Immunoaffinity Kit for IgG coupling trough Fc region    (pdf)
        BIO RAD   Protein A removal  from IgG on CHT Ceramic Hydroxyapatite Support   (pdf)
        BIOSEPRA Purification of Antibodies-Article  (pdf)
        BIOSEPRA  Validated Alternatives to Protein A Sorbents for Antibody Production.   (pdf)
        BIOSEPRA  MBI HYPERCELTM  Mixed-mode sorbent for direct capture of antibodies   (pdf)
        CALBIOCHEM   Human IgG subclasses   (pdf)   
        CLONTECH: Thiophilic - Purification of Immunoglobulins (pdf)  
        MERCK   Capture of Mouse Monoclonal Antibodies by Cation Exchange Chromatography   (pdf)   
        MILLIPORE   Affinity Chromatography Media: ProSep®-vA Ultra Media, ProSep-vA High Capacity Media, ProSep-A High Capacity Media &
                         ProSep-rA High Capacity Media. OPERATING INSTRUCTIONS ProSep-A affinity media have been developed specifically for industrial
                         scale purification of monoclonal antibodies where highly efficient purification can be achieved using clarified bioreactor feedstock at physiological
                         pH and salt concentration. (pdf)
        MILLIPORE  PROSEP-Thiosorb and PROSEP-Thiosorb M Chromatography Media - for the recovery and purification of Immunoglobulins   (pdf)
        PIERCE:  Binding Characteristics of Immunoglobulin Binding Proteins and Thiophilic Gel   (PowerPoint)
        PIERCE:  Immobilized E. coli Lysate. Partially purified bacterial proteins from a suspension of E. coli cells (strain BMH 71-18) immobilized onto
                        agarose, provides a simple and efficient method of removing E. coli-reactive proteins from antibody preparations   (pdf_I)   (pdf-II)
        PIERCE:  Antibody Immobilization: Choosing the Best Support   (PowerPoint)
        PIERCE:Immobilized Jacalin (human IgA and IgD binding lectin).
                        Jacalin immobilized on supports such as agarose has been useful for the purification of human serum or secratory IgA1    (pdf)    (pdf-II)
        PIERCE:  T-GelTM Purification Kit   (pdf)   
        SARTORIUS  Sartobind? Protein A 75 Membrane Adsorbers - Operating Instructions
                        A Separation Technology Based on Microporous Membranes (pdf)
       VIVASCIENCE: Vivapure Protein A - Mini Spin Columns (pdf)

General

        BIO RAD   Polymixin matrix - For removal of Endotoxin molecules (pdf)  
        MILLIPORE  Matrex Cellufine Sulfate - For concentration, purification and depyrogenation of Virus, Viral/Microbial Antigen,
                  Heparin binding proteins   (pdf)   
        PROMETIC BIOSCIENCES   Mimetic LigandTM Affinity Chromatography. The Mimetic Ligand adsorbent range has been designed to enable
                purification of many proteins, providing a general technique that can replace ion-exchange, gel permeation and hydrophobic interaction
                chromatography, but with higher recovery and purity. Mimetic Ligand Screening Column Kit contains 1ml pre-packed columns of ten different
                Mimetic LigandTM adsorbents, suitable for attachment to automated chromatography work stations. (pdf)
      
      Immunoprecipitation 免疫沉淀

          PIERCE:  SeizeTM X Immunoprecipitation Kits. Recover protein without antibody protein band interference. The Kit combine cross-linking
                and affinity chromatography to offer a new and improved immunoprecipitation method. First, the primary antibody is bound and immobilized to
                a Protein A or Protein G support using cross-linking agent (DSS). This properly orients the antibody to "seize" protein from crude cell lysate
                applied to the immobilized antibody support. Unbound proteins are then centrifuged away and the protein is recovered by using an elution buffer.
                (pdf -I)   (pdf-II)   (pdf-III)   (pdf-IV)

Lectins
and Glycoprotein Purification外源凝集素和糖蛋白纯化
   
       AMERSHAM-BIOSCIENCES  Con A Sepharose 4B  (Con A binds molcules containing Alpha-D-mannopyranosyl, Alpha-D-glucopyranosyl  and sterically related residues) (pdf)
       AMERSHAM-BIOSCIENCES  HiTrap Lectin Test Kit consists of four glycoprotein binding columns, HiTrap Con A, HiTrap Lentil Lectin,
                         HiTrap Wheat Germ Lectin and HiTrap Peanut Lectin.   (pdf)
       AMERSHAM-BIOSCIENCES   HiTrap Wheat Germ Lectin (high affinity to N-acetylglucosamine and reacts strongly with the chitobiose core of N-linked
                         oligosaccharides and N-acetylneuraminic acid)   (pdf)   
       AMERSHAM-BIOSCIENCES  Lentil Lectin Sepharose 4B (binds to polysaccharides and glycoconjugates containing glucose or mannose
                         type sugars)  (pdf)
        CALBIOCHEM   Lectins  [url=file:///C:/My%20Documents/site/PDF/affinity/CALBIOCHEM_Lectins.pdf] (pdf)[/url]PIERCE:  Immobilized Jacalin (human IgA and IgD binding lectin). Jacalin immobilized on supports such as agarose has been useful for the purification of
                         human serum or secratory IgA1    (pdf)    (pdf-II)   
        PIERCE:   Glycoprotein Isolation Kit, WGA isolates glycoproteins from complex protein mixtures using the lectin wheat germ agglutinin (WGA)
                         immobilized on agarose. The WGA lectin preferentially binds N-acetyl glucosamine (GlcNAC) and terminal GlcNAC structures that
                         are commonly present in many serum and membrane glycoproteins. WGA also has affinity for sialic acid.   (pdf)  
        PIERCE:   Glycoprotein Isolation Kit, ConA isolates glycoproteins from complex protein mixtures using the lectin concanavalin A (ConA) immobilized
                         on agarose. The ConA lectin preferentially recognizes α-linked mannose and, to a lesser extent, terminal glucose residues. These
                         carbohydrates   (pdf)  
        PROMETIC BIOSCIENCES   Aminophenyl Boronate Affinity Chromatography for capture of glycoproteins. The aminophenyl boronate-glycoprotein
                         covalent bond can be subsequently disrupted to elute the protein of interest with a pH change or by competitive elution using a sugar such as
                         sorbitol.   (pdf)
        QIAGEN:  Qproteome Glycoprotein Fractionation Handbook. For the fractionation of glycoproteins in proteomic samples   (pdf)
                          Qproteome Total Glycoprotein Kit. The Total Glycoprotein Spin Columns in the Qproteome Total Glycoprotein Kit contain ConA
                          and WGA lectins. They are used for a general enrichment of the total glycoprotein population from a cell or serum sample.
                          Qproteome Mannose Glycoprotein Kit. The ConA, GNA, and LCH lectin spin columns in the Qproteome Mannose Glycoprotein
                          Kit are used for specific enrichment of glycoproteins with mannose-rich glycan moieties. The three lectins each bind different
                          subclasses of these moieties.
                          Qproteome Sialic Glycoprotein Kit. The WGA, SNA, and MAL lectin spin columns in the Qproteome Sialic Glycoprotein Kit
                          are used for specific enrichment of glycoproteins with sialicacid-rich glycan moieties. The three lectins each bind different
                          subclasses of these moieties.
                          Qproteome O-Glycan Glycoprotein Kit. The AIL, and PNA lectin spin columns in the Qproteome O-Glycan Glycoprotein Kit are
                          used for specific enrichment of glycoproteins with a glycan structure of the type that are found on T-antigens. The two lectins each
                          bind different subclasses of these glycoproteins.
        QIAGEN:  Qproteome GlycoArrays is a simple, rapid, kit-based method for determining the pattern and relative abundance of specific
                          mammalian glycosylation epitopes in a glycosylated protein. The analysis can be performed on crude samples in growth media,
                          eliminating the need for time-consuming purification and sample preparation steps. The technology consists of arrays of selected lectins,
                          which are used to determine the glycosylation features of the analyzed glycoproteins.   (pdf)  

Albumin and Other Abundant Serum Proteins Removal        

AGILENTThe Agilent Multiple Affinity Removal System for Proteomics Sample Preparation. Designed to remove greater than 98-99%
                           of six interfering high-abundant proteins using Affinity-purified polyclonal antibodies to: albumin, IgG, IgA, transferrin, haptoglobin,
                           and antitrypsin from human serum or three proteins (albumin, IgG, and transferrin) from mouse serum samples   (pdf-I)   (pdf-II)
                            (pdf-III)  (pdf-IV)   (pdf-V)         AGILENT The Agilent Multiple Affinity Removal System. Designed to remove six interfering high-abundant proteins from Monkey
                           Plasma for Proteomics Sample Preparation   (pdf)
CALBIOCHEM   ProteoExtractTM Removal Kits (for Enhancing Resolution of Low Abundance Proteins)   (pdf)
        NORGEN   ProteoSpinTM Abundant Serum Protein Depletion Kit. The kit depletes 70% of albumin, 90% of α-antitrypsin, and 50% of transferin
                           and haptoglobin, enabling the visualization of low abundance proteins. The kit is unique in that it is based on an ion-exchange mechanism, silicon
                           carbide (SiC), and not the use of specific antibodies. As a result, the kit can be used to deplete serum proteins from a wide variety of samples,
                           including human and various animals. (pdf)
        PIERCE:     SwellGel® Blue Albumin Removal Kit   (pdf)  
        QIAGEN:   Qproteome Albumin/IgG Depletion Handbook. Fast and specific removal of albumin and IgG from human serum and plasma samples.
                           Based on monoclonal antibodies that bind HSA and human IgG with high affinity and specificity.   (pdf)
        SIGMA:      ProteoPrep 20 Plasma Immunodepletion Kit is a complete kit with all necessary reagents and consumable equipment to deplete 20
                           high abundance proteins from human plasma or serum. This kit is designed to specifically remove the 20 proteins from human plasma.
                           Specifically, 8 mL of plasma may be depleted in preparation for proteomic analysis, two-dimensional electrophoresis (2DE), or
                           liquid chromatography (LC).   (pdf).  
         VIVASCIENCE:  Vivapure® Anti-HSA Kit for Human Albumin Depletion
                          Highly specific human albumin depletion with unique antibody fragments   (pdf-I)   (pdf-II) (pdf-IV)   (pdf-V)
                          Vivapure Anti-HSA Affinity Resin   (pdf-III)   
         VIVASCIENCE:Vivapure® Anti-HSA/IgG Kits for Human Albumin and Human Albumin/IgG Depletion   (pdf )

Chromatofusing
   
      AMERSHAM-BIOSCIENCES  Chromatofusing Handbook(pdf)
      AMERSHAM-BIOSCIENCES    Ion Exchange Cromatography and Chromatofusing Handbook (pdf)

Crystallography and Recombinant Methods
     JENA Macromolecular crystallography products. Crystallization Screens: JBScreen Basic and JBScreen Membrane. Crystallization Optimization:
               JBS Solubility Kit, JBS Methylation Kit, JBScreen Plus, JBScreen Detergents, JBScreen Buffer Kits and JBScreen Cryo   (pdf)
      Derewenda Z.,  The use of recombinant methods and molecular engineering in protein crystallization. Methods (2004), 34: 354-363   (pdf)
      Smyth D.,  REVIEW  Crystal structures of fusion proteins with large-affinity tags.Protein Science (2003), 12:1313-1322 (pdf)   

Deglycosylation

      NORTHSTAR   Enzyme Deglycosylation Kit. Contains all enzymes needed to completely remove all N- & simple O-linked carbohydrates
                       from glycoproteins   (pdf)
      QAbio    Enzymatic CarboRelease Kit will remove all N-linked oligosaccharides and many O-linked oligosaccharides from glycoproteins. N-links
                     (Asn-linked) are removed using the enzyme PNGase F. In addition, all Ser/Thr-linked (O-linked) Gal-(β1-3)-GalNAc-(α1) and all sialic acid
                     substituted Gal-(β1-3)-GalNAc-(α1) will be removed using the combination of Sialidase and O-Glycosidase. The addition of ß-Galactosidase
                     and Hexosaminidase will assist in the deglycosylation of larger O-link structures. (pdf)
      PROZYME-GLYKO     Enzymatic Deglycosylation Kit. Contains all enzymes & reagents needed to completely remove all N-linked & simple
                      O-linked carbohydrates from glycoproteins   (pdf)
      ROCHE   The N-Glycosidase F Deglycosylation Kit can be used to test for the existence of asparagine-linked glycan chains on glycoproteins.
                      The new Endoglycosidase H Deglycosylation Kit is useful for testing the existence of "high mannose" type and "hybrid" type asparagine-linked
                      glycan chains on purified glycoproteins.   (pdf-I)    (pdf-II)
        SIGMA    Deglycosylation Selection Guide   (pdf)

Expanded Bed Adsorption

      AMERSHAM-BIOSCIENCESIntroduction to Expanded Bed Adsorption   (pdf)
       AMERSHAM-BIOSCIENCES Streamline Application Note  (pdf)
       AMERSHAM-BIOSCIENCES Streamline Data File (pdf)   

      BIORAD  CHT Ceramic Hydroxyapatite: Use in Expanded Bed Adsorption Mode   (pdf)
      BIOSEPRA Q & CM Ion Exchange Sorbents (pdf)  
     

Gel Filtration凝胶过滤纯化
    AMERSHAM-BIOSCIENCES-凝胶过滤手册Gel Filtration Handbook (pdf)  
      BIOSEPRA  Ultragel AcA - 分子筛层析Size Exclusion Chromatography Sorbents (pdf)   
      MERCK Fractogel® EMD BioSEC (S) (pdf)
      TOSOH BIOSCIENCE   分子筛层析Size Exclusion Chromatography   (pdf)

Hydrophobic Interaction Chromatography 亲水层析纯化

      AMERSHAM-BIOSCIENCES  Butyl-S SepharoseTM 6 Fast Flo   (pdf)       AMERSHAM-BIOSCIENCES  Hydrophobic Interaction and Reversed Phase Chromatography Principles and Methods   (pdf)
       AMERSHAM-BIOSCIENCES  Hydrophobic Interaction Chromatography Manual   (pdf)   
       AMERSHAM-BIOSCIENCES  HiTrapTM HIC Selection Kit consists of six Hydrophobic Interaction Chromatography media (HIC) with different
                        hydrophobic characteristics. The kit provides the possibility to screen for the most appropriate HIC medium for specific applications.
                           (pdf-I)   (pdf-II)
       AMERSHAM-BIOSCIENCES   RESOURCETM ETH (ether), ISO (isopropyl) and PHE (phenyl) are pre-packed high performance columns for
                        separating biomolecules by hydrophobic interaction chromatography (HIC)   (pdf)
       BIORAD  Macro-Prep Hydrophobic Interaction Chromatography Support - Instruction Manual   (pdf-I)   (pdf-II)
       MERCK  Cleaning and Regeneraton of  Fractogel EMD sorbents   (pdf)       MERCK  Fractogel EMDPhenyl (S) Hydrophobic Interaction Chromatography (HIC)   (pdf)
       MERCK  Fractogel EMD Propyl (S) Hydrophobic Interaction Chromatography(HIC)   (pdf)
       TOSOH BIOSCIENCE   Use of Hydrophobic Interaction Chromatography With a Non-Salt Buffer System for Improving Process
                        Economics in Purification of Monoclonal Antibodies (pdf)      
       TOSOH BIOSCIENCE   Hydrophobic Interaction Chromatography   (pdf)

Hydrophobic Charge-Induction Chromatography

       BIOSEPRA MEP HYPER CELL Product Information  (pdf)
       BIOSEPRA MEP HYPER CELL Product Note  (pdf)
       BIOSEPRA Purification of Antibodies-Article  (pdf)
       BIOSEPRA Validated Alternatives to Protein A Sorbents for Antibody Production.   (pdf)
       BIOSEPRA MBI HYPERCELTM  Mixed-mode sorbent for direct capture of antibodies   (pdf)

Hydroxyapatite

       BIORAD  Bio-Gel® HT - Bio-Gel HTP DNA Grade Bio - Gel HTP Hydroxyapatite - Instruction Manual   (pdf)
       BIORAD CHT Ceramic Hydroxyapatite   (pdf)
       BIORAD  CHT Ceramic Hydroxyapatite: Use in Expanded Bed Adsorption Mode   (pdf)
       BIORAD  CFT Ceramic Fluoroapatite - A Chromatographic Support  for Protein Purifications Requiring Acidic Conditions   (pdf)
       BIO RAD   Protein A removal  from IgG on CHT Ceramic Hydroxyapatite Support   (pdf)
       BIORAD   Macro Prep  Chromatography Supports   (pdf)
       BIOSEPRA   HA Ultragel Hydroxyapatite Chromatography Sorbents (pdf)

Ion Exchange Chromatography

       AMERSHAM-BIOSCIENCES   Ion Exchange Chromatography Manual   (pdf)  
        AMERSHAM-BIOSCIENCES    Ion Exchange Cromatography and Chromatofusing Handbook (pdf)   
        AMERSHAM-BIOSCIENCES    Ion Exchange Media - Selection Guide   (pdf)
        AMERSHAM BIOSCIENCES  Recommended buffers for Anion Exchange Chromatography
        AMERSHAM BIOSCIENCES  Recommended buffers for Cation Exchange Chromatography
        AMERSHAM BIOSCIENCES  Recommended Volatile buffers systems for Ion Exchange Chromatography
         AMERSHAM BIOSCIENCES   Capto adhere: a strong anion exchanger with multimodal functionality ionic interaction, hydrogen bonding and
             hydrophobic interaction. Capto adhere is designed for post Protein A purification of monoclonal antibodies. Removal of leached Protein A,
            aggregates, host cell proteins, nucleic acids and viruses from monoclonal antibodies is performed in flowthrough mode at where the antibodies
            pass directly through the column while the contaminants are adsorbed.   (pdf-I)   (pdf-II)   (pdf-III)
         AMERSHAM BIOSCIENCES   CaptoTM MMC: a multimodal cation exchanger. Has a multimodal ligand that may interact with target molecules
              in several different ways. It contains a carboxylic group and thus its features partly resemble those of a weak cation exchanger. In addition to
              the ionic interactions several other types of interactions are involved, including hydrogen bonding and hydrophobic interaction. The design of
              the ligand enables binding of proteins at high conductivity.  (pdf-I)  (pdf-II)  (pdf-III)
         AMERSHAM BIOSCIENCES   MacroCapTM SP is a highly porous cation exchanger high available surface area for adsorption of of large
            biomolecules  such as IgM and polyethylene glycol(PEG)-modified proteins (i.e., PEGylated proteins) that are intended for use as biopharmaceuticals.
               (pdf)
        AMERSHAM BIOSCIENCES   Q-Sepharose XL (anion exchange). Q-Sepharose XL virus licensed. SP-Sepharose XL (cation exchange) (pdf)   
        AMERSHAM BIOSCIENCES   CaptoTM Q a strong anion exchange medium for packed bed chromatography that allows increased speed and
                 throughput in capture and intermediate purification. It combines high capacity with high flow velocity and low backpressure (pdf)        BIORAD  Macro-Prep Ion Exchange Support - Instruction Manual  [url=file:///C:/My%20Documents/site/PDF/IonExchange/BIORAD_MacroPrep.pdf](pdf)[/url]
        BIORAD UNO TMQ&S Continuous    (pdf)  (pdf  II)
        BIOSEPRA Q, S, DEAE, CM Ceramic HyperD (pdf)
        MERCK  Cleaning and Regeneraton of   Fractogel EMD sorbents   (pdf)
        MERCK Ion Exchange Chromatography Using Fractogel EMD Tentacle Supports   (pdf)   
        MERCK Fractoprep® DEAE Weak Anion Exchange chromatography   (pdf)
        MERCK Fractoprep® SO3 - Strong Cation Exchange chromatography   (pdf)
        MERCK Fractoprep® TMAE  Strong Anion Exchange (pdf)
        MERCK Fractogel EMD COO- (S) (M) Weak Cation Exchange   (pdf)
        MERCK Fractogel EMD DEAE (S) (M) Weak Anion Exchange   (pdf)
        MERCK Fractogel EMD DMAE (S) (M) Weak Anion Exchange   (pdf)
        MERCK Fractogel EMD SE High Capacity (M) Strong Cation Exchange (pdf)
        MERCK Fractogel EMD SO3- (S) (M) Strong Cation Exchange   (pdf)
          MERCK Fractogel EMD TMAE (S) (M) Strong Anion Exchange   (pdf)
        MERCK Fractogel EMD TMAE High Capacity (M) Strong Anion Exchange   (pdf)   
        SARTORIUS  SartobindTM - Membrane Adsorbers Discs and Cassettes
                       Reusable Sartobind Membrane Ion Exchangers for Adjustable Filter Holders   (pdf)        SARTORIUS Sartobind® Membrane Adsorbers - A Separation Technology Based on Microporous Membrane Ion Exchangers   (pdf)
       SARTORIUS Sartobind? SingleSep Disposable Capsules - A Separation Technology Based on Microporous Membranes -Operating Instructions (pdf)
       TOSOH BIOSCIENCE  TSK-GEL BioAssyst Series Ion Exchange Column   (pdf)
       TOSOH BIOSCIENCE  Ion Exchange Chromatography   (pdf)   
       VIVASCIENCE:   VivapureTM IEX kits include everything required for rapid purification of protein samples or contaminant removal from protein
                     samples: clarification spin columns for initial sample clearing, Vivapure spin columns and ready-to-use buffers in different concentrations for
                     the protein bind-wash-elute steps, and VivaspinTM ultrafiltration devices for final sample concentration and desalting. Basic and acidic protein
                     purification kits.   (pdf)

Phospho-Protein Purification

       CLONTECH:  BD Phosphoprotein Kit User Manual (pdf) (pdf-II)
       QIAGEN  Phosphoprotein Purification Kit. For purification and analysis of phosphorylated proteins from eukaryotic cells     (pdf-I)   (pdf-II)

Polyubiquitin-modified proteins

        PIERCE   Ubiquitin Enrichment Kit. For the isolation and study of intracellular polyubiquitin-modified proteins. Through the use of a
                        high-binding affinity resin, polyubiquitinated proteins are isolated from cell or tissue lysates. The bound proteins can then be eluted
                        from the affinity resin and analyzed using the anti-ubiquitin antibody. (pdf)

Protein Aggregation

       INTEGRITY BIOSOLUTION   Protein Aggregation   (page)   
      
Eiler S., Overexpression, Purification, and Crystal Structure of Native ER alphaLBD  Protein Expression and Purification (2001) 22, 165-173 (pdf)
       PROTEIN EXPRESSION FACILITY OF THE HEBREW UNIVERSITY OF JERUSALEM   Heat shock growth procedure

Protein Labeling with Biotin

MOLECULAR PROBES    Biotin-XX Microscale Protein Labeling Kit.  This kit has been optimized for labeling  small amounts
               (20-100 μg) of purified proteins with molecular weights between 12 and 150 kDa, and contains everything needed
               to perform three labeling reactions and to separate the resulting conjugates from excess reactive biotin. (pdf)   
       MOLECULAR PROBES    FluoReporter ®Biotin Quantitation Assay Kit for biotinylated proteins. Sensitive fluorometric assay for
              accurately determining the number of biotin labels on a protein. The assay can detect from 4 to 80 pmol of biotin in a sample.   (pdf)

Protein Refolding/ Inclusion Bodies
     
       AMERSHAM-BIOSCIENCES  Purifying Challenging Proteins: Membrane proteins, Multiprotein complexes and Inclusion bodies   (pdf)
       AVIDIS-TECHNOLOGY Refolding Chromatography
             Refolding Chromatography with Mini-Chaperones (pdf)
       BIO-VECTRA Vectrase AT - Folding Proteins with Disulfide Bands
       BIO-VECTRA Vectrase CD - Quick Screening of  Refolding Conditions (pdf)
       BIO-VECTRA  Vectrase DK - Refolding at High Protein Concentration      
       BIO-VECTRA     Vectrase P - Protein Disulfide Isomerase (PDI) Mimic   (pdf-I) (pdf-II)
       GENO-Protein Foldase-Protein Folding Optimization Kit
       NORGEN  The ProteoSpinTM Inclusion Body Isolation Kit. Facilitates the isolation of recombinant proteins in the form of inclusion bodies from E.
            coli. The kit includes reagents specially formulated to achieve rapid and high-quality purification of inclusion body proteins using three processes:
                1. Lysis of bacterial cells to release inclusion bodies in solid form
                2. Solubilization of purified inclusion bodies
                3. Purification of the recombinant protein using spin column chromatography with Norgen's proprietary resin as an ion-exchanger
            Each spin column is able to purify up to 12 mg of recombinant proteins from 100 mL of culture. The kit is designed to purify both acidic and
            basic proteins.   (pdf Maxi Kit)   (pdf Micro Kit)
       NOVAGEN -Protein Refolding kit (pdf)
       NOVAGEN Information on Protein Refolding (pdf)
       NOVAGEN  iFOLDTM Protein Refolding System . The system includes inclusion body purification reagents combined with a dispensed 96-well
            plate-based protein refolding buffer matrix.   (pdf-I)    (pdf-II)
       NOVEXIN   The technology and reagents protect the protein during vulnerable steps in the refolding process, providing an opportunity for the
            protein to refold corrctly, reducing losses due to aggregation.
            Employs linear carbohydrate polymers of ~5kDa that enhance protein solubility and stability through the formation of reversible complexes with
            proteins without altering their structure and preventing aggregation.
            Protein Refolding  Starter Kit   (pdf)
            Stability P.A.C.   (pdf)
            Acidic Protein Refolding Kit   (pdf)
            Basic Protein Refolding Kit   (pdf)  
            Refolding Kit for Histidine-tagged proteins   (pdf-I)   (pdf-II)
            Bulk Protection and Release Reagents   (pdf)
       PIERCE   Pro-MatrixTM Protein Refolding Kit (pdf)       Additives Used to Increase Folding and Prevent Aggregation
PROTEIN EXPRESSION FACILITY OF THE HEBREW UNIVERSITY OF JERUSALEM   Heat shock growth procedure
       PROTEIN EXPRESSION AND PURIFICATION FACILITY OF THE EUROPEAN MOLECULAR BIOLOGY LABORATORY
             In vitro denaturation and refolding       Contaminant Removal from Inclusion Bodies Before Solubilization
      UNIVERSITY OF OKLAHOMA  School of Chemical Engineering and Materials Science Recombinant Protein Solubility Prediction

    Recommended Reading

     Altamirano M., Refolding Chromatography with Immobilized Mini-Chaperones. PNAS 1997, 94: 3576-3578 (pdf)       Armstrong N., A New Protein Folding Screen...etc. Protein Science 1999,  8: 1475-1483   (pdf)
       Chen G., Overexpression of a Glutamate Receptor (GluR2) ligand binding domain in E.Coli: Application of a novel protein folding screen   (pdf)
       De Bernardez Clark E.,Refolding of  RecombinantProteins  . Current Opinion in Biotechnology 1998,9:157-163   (pdf)
       De Bernardez Clark E., Protein refolding for industrial processes. Current Opinion in Biotechnology 2001, 12:202-207 (pdf).
       Eiler S., Overexpression, Purification, and Crystal Structure of Native ER alphaLBD  Protein Expression and Purification (2001) 22, 165-173   (pdf)       Machida S.,  Cycloamylose as an efficient artificial chaperone for protein refolding.  FEBS Letters 486 (2000) 131-135 (pdf)       Middelberg A., Preparative Protein Folding. TRENDS in Biotechnology 2002, 20 (10): 437-443 [url=file:///C:/My%20Documents/site/PDF/Literature/Middelberg2002.pdf] [/url] (pdf)
       Ming Li et al., In vitro protein refolding by chromatographic procedures. Protein Expr. and Purif. 2004,33: 1-10   (pdf)
       Tsumoto K., Practical Considerations in Refolding Proteins from Inclusion Bodies. Protein Expr. and Purif. 2003,28: 1-8
(pdf)      

  Reverse Phase Chromatography
   
        AMERSHAM-BIOSCIENCES  Hydrophobic Interaction and Reversed Phase Chromatography Principles and Methods   (pdf)
        AMERSHAM-BIOSCIENCE  Reverse Phase Chromatography Manual   (pdf)
        VYDAC   The Handbook of Analysis and Purification of Peptides and Proteins by Reverse-Phase HPLC   (pdf)    

Storage of Purified Proteins
   
       PROTEIN EXPRESSION AND PURIFICATION FACILITY OF THE EUROPEAN MOLECULAR BIOLOGY LABORATORY                   Storage of Purified Proteins       PIERCEProtein stability and storage   (pdf)

Thiophilic Chromatography
    CLONTECH: Thiophilic - Purification of Immunoglobulins (pdf)  
      CLONTECH:   Protein Purification Products (pdf)
       AMERSHAM-BIOSCIENCE   Activated Thiol Sepharose 4B reacts with solutes containing thiol groups under mild conditions to form mixed disulphides.
            This reaction forms the basis of covalent chromatography and a procedure for immobilizing thiol containing biomolecules.   (pdf)
      AMERSHAM-BIOSCIENCES Thiopropyl Sepharose  (pdf)
      MERCK  Fractogel Thiophilic (pdf)
      PIERCE:  T-GelTM Purification Kit   (pdf)      
      MILLIPORE PROSEP-Thiosorb and PROSEP-Thiosorb M Chromatography Media - for the recovery and purification of Immunoglobulins   (pdf)

Viral Purification

    AMERSHAM BIOSCIENCES   Q-Spharose XL (anion exchange). Q-Spharose XL virus licensed. SP-Spharose XL (cation exchange)  (pdf)
    AMERSHAM BIOSCIENCES   Rapid Adenovirus Purification Using Q-Spharose XL (pdf)     CLONTECH: Adeno-XTM Virus Purification Kits User Manual   (pdf-I)  Protocol (pdf-II). A complete filtration-based system for purifying and
            concentrating recombinant adenovirus. It provides a superior alternative to cesium chloride (CsCl) density gradient centrifugation. Use an
            adsorbent membrane that selectively binds adenoviral particles based on their distinctive surface-associated properties.
    MILLIPORE  Matrex Cellufine Sulfate - For concentration, purification and depyrogenation of Virus, Viral/Microbial Antigen, Heparin binding proteins   (pdf)
    PALL   Dynamic High Capacity Mustang® Q Membrane Units for Scaleable Anion Exchange Chromatography Purification of Adenoviral Vectors   (pdf)
   VIVASCIENCE:  Vivapure® AdenoPACKTM 100. Adenovirus (Ad5) purification and concentration kit for up to 100 ml cell culture volume.
            AdenoPACK syringe filters containing an ion exchange membrane adsorber that binds adenoviral particles. (pdf-I)   (pdf-II)  (pdf-III)      

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