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Protein Expression & Purification


Products >  Protein_Expression_and_Purification >  GST-Tagged_Protein_Purification >  GST_Purification_Resin

GST-Tagged Protein Purification Kit

Glutathione-Superflow allows rapid affinity purification of GST-tagged proteins for batch, gravity flow, and FPLC applications. The resin is based on 6% cross-linked agarose, with glutathione covalently bound to the resin. It can withstand flow rates as high as 15 ml/cm2/min and medium pressure (up to 150 psi).

The GST Purification Kit contains sufficient buffers and Glutathione-Superflow Columns for performing five batch/gravity-flow purifications. Up to 10 mg per prep of GST-tagged recombinant proteins can be purified using this kit.

GST Tags

Epitope tags such as glutathione-S-transferase (GST) are often used as tags for proteins for expression and purification applications. Glutathione transferases are abundant enzymes involved in cellular defense against electrophilic chemical compounds, which bind glutathione with high affinity and specificity. The strength and selectivity of this interaction can be exploited to effectively purify GST-tagged proteins. Glutathione-Superflow Resin selectively binds the GST-tagged protein under normal conditions, allowing the protein of interest to be separated from whole cell extracts rapidly and efficiently. A high degree of purification can be achieved in just one chromatographic step.

GST is a 35 kDa protein that can be assayed biochemically as well as immunologically. This characteristic sets it apart from many epitope tags that are simply short peptides. However, since GST is a large tag, the potential for degradation by proteases is higher than for smaller tags. Therefore, performing GST-based protein purification as quickly as possible under nondegrading conditions is necessary to minimize sample loss. GST loses its ability to bind Glutathione-Superflow Resin when denatured, and consequently strong denaturants such as guanidine-HCl or urea must not be used in the purification buffers. Check reagent compatibilities when designing your purification scheme.

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Features

  • Easy to use—no optimization required
  • Purifies more protein—high binding capacity (>10 mg of recombinant GST fusion protein per ml resin)
  • Yields highly purified protein—eluted protein is free of cellular contaminants
  • Reusable—easy protocol for regenerating and storing resin
  • Save money—30–40% less expensive than competing brands

Applications

  • Purification of GST-tagged proteins: batch, gravity flow, or FPLC applications

Reagents Compatible with Glutathione-Superflow Resin

Reagent Acceptable Concentration
Beta-mercaptoethanol 10 mM (with caution)
EDTA, EGTA 10 mM
Ethanol 30%, for regeneration only
Glycerol 20%
HEPES 50 mM
Imidazole 200 mM at pH 7.0–8.0
KCl 500 mM
MES 20 mM
MOPS 50 mM
NaCl 1.0 M
NP-40 1%
Tris 50 mM

Reagents Incompatible with Glutathione-Superflow Resin

These reagents are incompatible at any concentration:

  • CHAPS
  • DTE (dithioerythritol) and DTT (dithiothreitol)
  • Guanidine hydrochloride
  • SDS
  • Urea

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


 
 
Products
Cat. # Product Contents Size Price Units Select
635619 GST Purification Kit 5 Purifications $370.00
635607 Glutathione-Superflow Resin 10 mL $190.00
635608 Glutathione-Superflow Resin 100 mL $1,441.00
 

 

 
 

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