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GE17-5130-01 Sigma

GSTrap Fast Flow

GE Healthcare, 17-5130-01, pack of 5 × 1 mL

  •  NACRES NA.25

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Properties

Related Categories Affinity Chromatography, Glutathione, Molecular Biology, Protein Chromatography, Proteomics More...
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 5 × 1 mL
mfr. no.   GE Healthcare, 17-5130-01
parameter   flow rate
  42 psi
bed size   7 mm × 25 mm
bed volume   1 mL
column I.D.   7 mm
matrix   highly cross-linked 4% agarose
average diameter   90 μm
cleaning   3 - 12
working range   3 - 12
capacity   >10 mg/mL binding capacity(The pressure over the packed bed varies depending on a range of parameters such as the characteristics of the chromatography medium and the column tubing used.)

Description

General description

GSTrap FF columns are prepacked Glutathione Sepharose® Fast Flow columns for fast, convenient, one-step purification of glutathione S-transferase (GST) tagged proteins.

GST*-tagged proteins can be purified directly from pretreated bacterial lysates using GSTrap FF.GST-tagged proteins are eluted under mild, nondenaturing conditions using reduced glutathione. The purification process preserves protein antigenicity and function. If desired, cleavage of the protein from GST can be achieved using a site-specific protease whose recognition sequence is located immediately upstream from the multiple cloning site on the pGEX plasmids. GST-tagged proteins can be detected using colorimetric or immunological methods. The medium, Glutathione Sepharose® 4 Fast Flow, is also available as lab packages and is an excellent choice for scale-up.The columns can be operated with a syringe, peristaltic pump, or liquid chromatography system such as AKTA design or FPLC System.

* See the section for licensing information in our terms and conditions.

Features and Benefits

• Fast and simple one-step purification of GST-tagged proteins.
• Mild elution conditions preserving protein antigenicity and function.
• Easy scale-up by connecting the columns in series.
• Sepharose® 4 Fast Flow provides good flow properties.

Analysis Note

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

Legal Information

FPLC is a trademark of GE Healthcare

GSTrap is a trademark of GE Healthcare

Sepharose is a registered trademark of GE Healthcare

Safety & Documentation

Safety Information

Symbol 
GHS02  GHS02
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
UN1170 - class 3 - PG 3 - Ethanol, solution

Documents

Certificate of Analysis

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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 GSTrap

The protocol below is an example optimized for 8 mg of target protein. It is worth estimating how much target protein is applied to the column, as this allows one to minimize the amount of protease a...
Keywords: Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Digestions, Filtration, PAGE, Purification, Sonication, Tagged proteins

Cleavage and Purification of GST-Tagged Protein Eluted from GSTrap

The protocol below is an example optimized for 8 mg of target protein. It is worth estimating how much target protein is applied to the column, as this allows one to minimize the amount of protease a...
Keywords: Absorption, Buffers, Chromatography, Digestions, Purification

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

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

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

Manual and Automated Purification

Chapter 2, Extracted from Affinity Chromatography Vol. 2: Tagged Proteins, GE Healthcare, 2016
Keywords: Affinity chromatography, Centrifugation, Chromatography, Gene expression, 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

GSTPrep FF 16/10 columns are based on the 20 ml HiPrep column design, ready to use for easy, one-step preparative purification of GST-tagged proteins, other glutathione S-transferases, and glutathion...
Keywords: Addition reactions, Buffers, Centrifugation, Chromatography, Homogenization, Oxidations, PAGE, Purification, Sample preparations, Sonication, Tagged proteins

Preparative Purification using GSTPrep FF 16/10 column

GSTPrep™ FF 16/10 columns are based on the 20 ml HiPrep column design, ready to use for easy, one-step preparative purification of GST-tagged proteins, other glutathione S-transferases, and glutathion...
Keywords: Addition reactions, Affinity chromatography, Buffers, Centrifugation, Chromatography, Homogenization, Oxidations, PAGE, Sample preparations, Sonication, Tagged proteins

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 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

Removal of Thrombin and Factor Xa Using HiTrap Benzamidine FF (High Sub)

To protect the fusion protein from proteolytic degradation prior to enzymatic cleavage with PreScission Protease, thrombin, or Factor Xa, it may be necessary to remove proteases from the sample. Addi...
Keywords: Absorption, Cell culture, Chromatography, Degradations, PAGE, Purification

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

Selecting Equipment for Purification of GST-tagged Proteins

The choice of equipment is dependent on the specific purification. Many purification procedures can be carried out using simple equipment and methods, for example, in a step-gradient elution method u...
Keywords: Chromatography, Purification

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

Simple Purification Using a GSTrap Column with ÄKTAprime™

ÄKTAprime, in combination with pre-installed method templates for purifications and prepacked columns, is designed to perform the most common protein purification steps with the touch of one button. ...
Keywords: Affinity chromatography, Buffers, Cell disruption, Centrifugation, Chromatography, Filtration, PAGE, Purification, Size-exclusion chromatography, Sonication, 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

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