G4510
Glutathione−Agarose
lyophilized powder
Synonym(s):
Glutathione-Agarose Resin, GSH-agarose, S-linked glutathione agarose
About This Item
form
lyophilized powder
Quality Level
analyte chemical class(es)
proteins (GST)
extent of labeling
≥10 μmol per mL gel
technique(s)
immunoprecipitation (IP): suitable
protein purification: suitable
matrix
4% cross-linked beaded agarose
matrix activation
epoxy
matrix attachment
sulfur
matrix spacer
12 atoms (10 carbon)
storage temp.
−20°C
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General description
Application
- to purify glutathione S-transferase (GST)-fused protein for generating polyclonal antibody M4P
- to incubate supernatant fractions for the purification of full-length p53 (aa 1-393) and its DNA-binding domain (aa 94-288)
- in affinity purification of the fusion protein
Biochem/physiol Actions
Features and Benefits
- Offers rapid, mild, non-denaturing, and highly selective purification of glutathione-binding enzymes.
- Has a shelf life of two years if stored properly desiccated at -20 °C.
- Binding capacity: 5-10 mg glutathione S-transferase per mL resin
Quantity
Physical form
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
Regulatory Information
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Find documentation for the products that you have recently purchased in the Document Library.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How much Product G4510, Glutathione-Agarose, do I receive and how do I swell my powder?
This material is sold by the final volume of gel produced after swelling of the powder. One gram of powder swells to approximately 14 mL of gel. (Approximately 70 mg of powder swells to 1 mL of gel.) Swell the lyophilized powder in deionized water (use of some buffers can result in non-recoverable loss of volume) at 200 mL/g. Typically 90 to 95% of swelling occurs within 30 minutes at room temperature, but it may require overnight at 2 to 8 °C for 100% swelling to occur. After swelling, the agarose beads must be washed thoroughly with 10 volumes of water or equilibration buffer (such as PBS) to remove the lactose present in the lyophilized product.
How is Product G4510, Glutathione-Agarose, resin conjugated to the glutathione, and what is the binding capacity of the resin?
This product consists of glutathione attached through the sulfur to epoxy activated 4% cross-linked beaded agarose, resulting in a 12 atom spacer. Binding capacity: approx. 5 to 10 mg glutathione S-transferase per mL resin.
When using Product G4510, Glutathione-Agarose, how do I store the swollen resin?
After the resin has been swollen, it can be stored in 2 M NaCl at 2 to 8 °C. For long-term storage, a bacteriostatic agent such as 1 mM sodium azide or 0.02% thimerosal should be included. Alternatively, the resin can be relyophilized after addition of lactose.
How do I regenerate Product G4510, Glutathione-Agarose, resin?
Regeneration method:1. Wash with approximately 5 resin volumes of 0.1 M borate buffer, pH 8.5 containing 0.5 M NaCl (Cleansing Buffer 1).2. Wash with at least 5 column volumes of deionized water.3. Wash with at least 5 column volumes of 0.1 M acetate buffer, pH 4.5, containing 0.5 M NaCl (Cleansing Buffer 2).4. Wash with approximately 5 resin volumes of deionized water. For long-term storage, store in 2 M NaCl containing a bacteriostatic agent.5. Equilibrate with equilibration buffer before use.
What reagents are compatible with Product G4510, Glutathione-Agarose, resin?
Tris or phosphate buffers (pH 6.5 to 9.5) are typical lysis buffers compatible with glutathione affinity chromatography. Salt concentrations of up to 1 Molar do not interfere with binding. Protease inhibitors such as EDTA (Product No. E7889) or PMSF (Product No. P7626) are often included in the lysis buffer. Serine protease inhibitors included in the lysis buffer will not interfere with subsequent thrombin or factor Xa treatment as these inhibitors are removed before the proteolysis step. The binding of GST to glutathione-agarose is unaffected by 1% Triton X-100, 1% Tween-20, 1% CTAB, 10 mM DTT or 0.03% SDS.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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