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GE17-0756-04 Sigma

Glutathione Sepharose® 4B

GE Healthcare, 17-0756-04, pack of 300 mL

  •  NACRES NA.56

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Properties

Related Categories Affinity Chromatography, Avidin Biotin Matrices, Glutathione, Molecular Biology, Protein Chromatography,
shelf life   Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.
packaging   pack of 300 mL
mfr. no.   GE Healthcare, 17-0756-04
matrix   4% agarose
particle size   45-165 μm
average diameter   90 μm
cleaning in place   4 - 13
working range   4 - 13
capacity   >25 mg binding capacity (horse liver GST/ml medium)(Binding of GST to glutathione is flow-rate dependent and lower flow rates often increase the binding capacity. This is important to consider during sample loading and elution.)

Description

General description

Glutathione Sepharose® 4B is designed for high capacity single-step purification of Glutathione S-Transferase (GST) tagged fusion proteins.

GST-tagged proteins and other glutathione S-transferases or glutathone-dependent proteins can be purified directly from bacterial lysates using Glutathione Sepharose® 4B. Proteins are eluted under mild, non-denaturing conditions that preserve protein antigenicity.

Features and Benefits

• High capacity single step purification of recombinant glutathione S-transferase (GST) tagged proteins
• Proteins are eluted under mild, non-denaturing conditions that preserve protein antigenicity and functionality.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.gelifesciences.com.

Legal Information

Sepharose is a registered trademark of GE Healthcare

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Protocols

Affinity Chromatography Troubleshooting

This section focuses on practical problems that may occur when running a chromatography column. The diagrams below give an indication of how a chromatogram may deviate from the ideal during affnity p...
Keywords: Adsorption, Affinity chromatography, Buffers, Chromatography, Detergents, Digestions, Fractionation, Ligands, PAGE, Precipitation, Purification, Sample preparations, Separation, Solvents

Buffer Exchange and Desalting for Affinity Chromatography

Dialysis is frequently mentioned in the literature as a technique to remove salt or other small molecules and to exchange the buffer composition of a sample. However, dialysis is generally a very slo...
Keywords: Absorption, Affinity chromatography, Buffers, Chromatography, Dialysis, Separation

Characteristics of Glutathione Sepharose Products

Appendix 2, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016  
Keywords: Affinity chromatography, Buffers, Centrifugation, Chromatography, Inductively coupled plasma, Ligands, Purification, Tagged proteins

Characteristics of Glutathione Sepharose and HiTrap Benzamidine FF (High Sub) Media and Columns

Glutathione Sepharose High Performance is recommended for high-resolution purification of GST-tagged proteins, providing sharp peaks and concentrated eluent. Glutathione Sepharose 4 Fast Flow is exce...
Keywords: Buffers, Centrifugation, Chromatography, Inductively coupled plasma, Ligands, Purification, Tagged proteins

Cleavage and Purification of GST-Tagged Protein Bound to Glutathione Sepharose in Batch Mode

Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, and Glutathione Sepharose 4B can all be used for cleavage and purification of GST-tagged proteins in batch.
Keywords: Buffers, Cell disruption, Centrifugation, Chromatography, Purification, Tagged proteins

Column Packing and Preparation for Affinity Chromatography of GST-tagged Proteins

This appendix provides column packing guidelines for affinity chromatography media. Efficient column packing is essential for a good separation, especially when using gradient elution.
Keywords: Affinity chromatography, Buffers, Chromatography, Purification, Separation

Column Packing and Preparation for Affinity Chromatography

Appendix 3,  Extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007
Keywords: Affinity chromatography, Antimicrobials, Buffers, Chromatography, Separation

Column Packing and Preparation

Appendix 6, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016  
Keywords: Affinity chromatography, Antimicrobials, Chromatography, Gene expression, Separation, Tagged proteins

Desalting/Buffer Exchange and Concentration for Affinity Chromatography of Tagged Proteins

Desalting at laboratory scale is a well-proven, simple, and very fast method that will rapidly remove low molecular weight contaminants at the same time as transferring the sample into the desired bu...
Keywords: Absorption, Addition reactions, Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Dialysis, Fractionation, Ion Exchange, PAGE, Precipitation, Sample preparations, Separation, Size-exclusion chromatography, Tagged proteins

GST Detection Module with CDNB Enzymatic Assay

GST-tagged proteins produced using pGEX vectors can be detected enzymatically using the GST substrate CDNB, included in the GST Detection Module. The GST-mediated reaction of CDNB with glutathione pr...
Keywords: Cell disruption, Tagged proteins

General Considerations for Purification of GST-tagged Protein

The yield of GST-tagged proteins is highly variable, ranging from 1 mg/l to 10 mg/l. The yield depends on various parameters, such as nature of the tagged protein, the host cell, and the culture cond...
Keywords: Cell culture, Cell disruption, Detection methods, Dialysis, Filtration, Homogenization, PAGE, Purification, Sonication, Tagged proteins, Western blot

General considerations for purification of GST-tagged proteins

Yield of tagged protein is highly variable and is affected by the nature of the tagged protein, the host cell, and the expression and purification conditions used. Tagged protein yields can range from...
Keywords: Affinity chromatography, Cell disruption, Chromatography, Detection methods, Dialysis, Gene expression, Homogenization, PAGE, Sonication, Tagged proteins, Western blot

Gravity-Flow Purification Using GST GraviTrap

GST GraviTrap™ is designed for fast and simple purification of GST-tagged proteins using gravity flow. The column is prepacked with 2 ml of Glutathione Sepharose® 4B, enough to purify up to 50 mg of rG...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell culture, Chromatography, Homogenization, Oxidations, Sample preparations, Sonication, Tagged proteins

Gravity-flow Purification Using GST GraviTrap

GST GraviTrap is designed for fast and simple purification of GST-tagged proteins using gravity flow. The column is prepacked with 2 ml of Glutathione Sepharose 4B, enough to purify up to 50 mg of GS...
Keywords: Buffers, Cell culture, Homogenization, Oxidations, PAGE, Purification, Sample preparations, Sonication, Tagged proteins

High-throughput Screening Using GST MultiTrap FF and GST MultiTrap 4B 96-well Filter Plates

GST MultiTrap FF and GST MultiTrap 4B (Fig 3.4) are prepacked, disposable 96-well filter plates for reproducible, high-throughput screening of GST-tagged proteins. The plates are filled with a define...
Keywords: Buffers, Cell disruption, Centrifugation, Filtration, Gene expression, Oxidations, PAGE, Purification, Sample preparations, Sonication, Tagged proteins

High-throughput screening using GST MultiTrap 4B 96-well filter plates

GST MultiTrap™ 4B (Fig 5.4) is a prepacked, disposable 96-well filter plates for reproducible, high-throughput screening of GST-tagged proteins. Typical applications include expression screening of di...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Gene expression, Oxidations, Sample preparations, Tagged proteins

Minipreps Using GST SpinTrap

GST SpinTrap™ columns are designed for rapid, small-scale purification of GST-tagged proteins using conditions of mild affinity purification. Greater than 90% purity can be achieved in a single step. Th...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell culture, Cell disruption, Centrifugation, Chromatography, Gene expression, Homogenization, Oxidations, Protein extraction, Sample preparations, Sonication, Tagged proteins

Performing a Separation with GE Healthcare Products Based on GST Fusion Proteins

Glutathione S-transferase (GST) is one of the most common tags used to facilitate the purifcation and detection of recombinant proteins and a range of products for simple, one step purifcation of GST...
Keywords: Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Ligands, PAGE, Purification, Tagged proteins

Preparative Purification Using GSTPrep FF 16/10 Column

Glutathione Sepharose 4B, FF, and HP are available in lab packs, varying from 10 to 500 ml, for affinity purification of GST-tagged proteins using batch- or column-based methods. Glutathione Sepharos...
Keywords: Addition reactions, Buffers, Cell disruption, Centrifugation, Chromatography, Column chromatography, Gene expression, Homogenization, Oxidations, PAGE, Purification, Sample preparations, Sonication, Tagged proteins

Pull-down assays

Extracted from Purifying Challenging Proteins - Principles and Methods, GE Healthcare, 2007
Keywords: Buffers, Cell disruption, Centrifugation, Chromatography, Cloning, Digestions, Gene expression, Genetic, Immobilization, Immunoprecipitation, Ligands, Mass spectrometry, PAGE, Precipitation

Purifcation or Removal of Serine Proteases, e.g. Thrombin and Trypsin, and Zymogens

Sample extraction procedures often release proteases into solution, requiring the addition of protease inhibitors to prevent unwanted proteolysis. An alternative to the addition of inhibitors is to u...
Keywords: Absorption, Affinity chromatography, Buffers, Cell culture, Cell disruption, Chromatography, Detergents, Ligands, PAGE

Purification Options for GST-tagged proteins using Glutathione Sepharose Products

Table 5.1 Purification options for GST-tagged proteins using Glutathione Sepharose Products
Keywords: Affinity chromatography, Chromatography, Purification, Tagged proteins

Purification Using GST HiTrap 1 ml and 5 ml Columns: GSTrap 4B, GSTrap FF, and GSTrap HP

GSTrap affinity columns are specially designed 1 ml and 5 ml HiTrap columns packed with Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, or Glutathione Sepharose 4B media. R...
Keywords: Buffers, Chromatography, Oxidations, PAGE, Purification, Tagged proteins, Western blot

Purification Using GST SpinTrap

GST SpinTrap columns are designed for rapid, small-scale purification of GST-tagged proteins using mild conditions of affinity purification. More than 90% purity can be achieved in a single step. The...
Keywords: Addition reactions, Buffers, Cell culture, Cell disruption, Centrifugation, Gene expression, Homogenization, Oxidations, PAGE, Protein extraction, Purification, Sample preparations, Sonication, Tagged proteins

Purification using Glutathione Sepharose High Performance, Glutathione Sepharose 4 Fast Flow, and Glutathione Sepharose 4B

These three chromatography media are all used for the purification of GST-tagged recombinant proteins and other S-transferases or glutathione-dependent proteins. They allow mild elution conditions tha...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Homogenization, Ligands, Oxidations, PAGE, Sample preparations, Sonication, Tagged proteins

Purification Using GSTrap HP, GSTrap FF, and GSTrap 4B columns

GSTrap™ affinity columns are specially designed 1 ml and 5 ml HiTrap columns packed with Glutathione Sepharose® High Performance, Glutathione Sepharose® 4 Fast Flow, or Glutathione Sepharose® 4B chrom...
Keywords: Addition reactions, Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Homogenization, Oxidations, PAGE, Purification, Sample preparations, Size-exclusion chromatography, Sonication, Tagged proteins, Western blot

Removal of GST tag by enzymatic cleavage

Removal of the GST tag is often necessary to be able to perform functional or structural studies of the target protein. Tagged proteins containing a PreScission Protease, thrombin, or Factor Xa recog...
Keywords: Absorption, Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Digestions, PAGE, Size-exclusion chromatography, Sonication, Tagged proteins

Sample Preparation for Affinity Chromatography

Appendix 1 extracted from Affinity Chromatography Principles and Methods, GE Healthcare, 2007
Keywords: Adsorption, Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Filtration, Nucleic acid denaturation, PAGE, Precipitation, Sample preparations

Screening pGEX Recombinants for Tagged Protein Expression

Sections of this procedure have been adapted with permission from Current Protocols in Molecular Biology, Vol. 2, Supplement 10, Unit 16.7. Copyright © 1990 by Current Protocols.
Keywords: Buffers, Cell disruption, Gene expression, Molecular biology, PAGE, Protein extraction, Purification, Sonication, Sterilizations, Tagged proteins

Selecting a GST Affinity Chromatography Product

A variety of affinity chromatography products are available from GE Healthcare that have glutathione immobilized as ligand: Sepharose High Performance (HP), Sepharose 4 Fast Flow (FF), and Sepharose ...
Keywords: Affinity chromatography, Cell disruption, Chromatography, Immobilization, Ligands, Purification, Tagged proteins

Starting materials needed for the purification of integral membrane proteins for structural and functional studies

Extracted from Purifying Challenging Proteins - Principles and Methods, GE Healthcare, 2007
Keywords: Cloning, Detergents, Gene expression, Glycosylations, PAGE, Positron Emission Tomography, Separation, Tagged proteins

Troubleshooting Cleavage Methods

The troubleshooting guide below addresses problems common to the majority of cleavage methods as well as problems specific to a particular method. In the latter case, the relevant method is indicated....
Keywords: Affinity chromatography, Chromatography, Cloning, Electrophoresis, Tagged proteins, Western blot

Troubleshooting Purification Methods

The troubleshooting guide below addresses problems common to the majority of purification methods as well as problems specific to a particular method. In the latter case, the relevant method is indicat...
Keywords: Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Degradations, Detergents, Electrophoresis, Gene expression, Ion Exchange, Precipitation, Sonication, Tagged proteins, Western blot

Troubleshooting Strategies in Purification of GST-tagged Proteins

This troubleshooting guide addresses the common problems associated with the majority of purification methods using the different Glutathione Sepharose media.
Keywords: Buffers, Cell disruption, Centrifugation, Degradations, Detergents, Electrophoresis, Gene expression, Ion Exchange, Precipitation, Purification, Sonication, Tagged proteins, Western blot

Troubleshooting of Cleavage Methods

The troubleshooting guide below addresses problems common to the majority of cleavage methods as well as problems specific to a particular method. In the latter case, the relevant method is indicated.
Keywords: Cloning, Electrophoresis, Purification, Tagged proteins, Western blot

Troubleshooting of Problems Associated with the Expression and Growth of GST-tagged Proteins

The troubleshooting guide below addresses common problems associated with the expression and growth of GST-tagged proteins.
Keywords: Cell disruption, Gene expression, PAGE, Purification, Sonication, Tagged proteins, Western blot

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