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G4510
Glutathione−Agarose
lyophilized powder
Glutathione−Agarose
Synonym(s):
Glutathione-Agarose Resin, GSH-agarose, S-linked glutathione agarose
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1 ML
$115.00
5 ML
$362.00
10 ML
$496.00
50 ML
$1,780.00
About This Item
Form:
lyophilized powder
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form
lyophilized powder
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
Quality Level
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This Item | A8580 | SAB4200692 | A6306 |
|---|---|---|---|
| technique(s) immunoprecipitation (IP): suitable, protein purification: suitable | technique(s) ELISA: suitable, immunoprecipitation (IP): suitable | technique(s) ELISA: suitable, dot blot: suitable, immunoblotting: 0.5 μg/mL, immunofluorescence: suitable, immunoprecipitation (IP): suitable | technique(s) - |
| form lyophilized powder | form PBS suspension | form buffered aqueous solution | form lyophilized powder |
| storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. 2-8°C |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| matrix 4% cross-linked beaded agarose | matrix - | matrix - | matrix - |
| analyte chemical class(es) proteins (GST) | analyte chemical class(es) - | analyte chemical class(es) - | analyte chemical class(es) - |
General description
Glutathione S-transferase (GST) is a protein with a molecular weight of 26 KDa that naturally occurs in eukaryotic cells. It belongs to the family of Phase II detoxification enzymes. Glutathione-Agarose consists of glutathione attached through its sulfur to epoxy-activated 4% cross-linked beaded agarose, resulting in a 12-atom (10-carbon) spacer. It is provided as a lyophilized powder stabilized with lactose.
Application
Affinity chromatography using glutathione-agarose permits rapid, mild, non-denaturing and highly selective purification of proteins containing glutathione binding sequences, such as Glutathione S-Transferase (GST), glutathione peroxidase and glyoxalase I.
Glutathione-Agarose has been used:
- 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
Glutathione S-transferases (GSTs), a superfamily of multifunctional detoxification isoenzymes, have a significant role in cellular signaling. GSTs are emerging as promising therapeutic targets due to the overexpression of specific isozymes in various tumors and their potential involvement in the development of other diseases such as neurodegenerative diseases, multiple sclerosis, and asthma.
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
Physical form
Lyophilized powder stabilized with lactose
Preparation Note
One g powder swells to ≥10 ml of gel
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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Related Content
Investigate in vitro protein-protein interactions with pull-down assays, utilizing affinity, GST pull-down, TAP, and co-immunoprecipitation methods.
Investigue las interacciones proteína-proteína in vitro con análisis pull-down, utilizando métodos de afinidad, pull-down con GST, TAP y co-inmunoprecipitación.
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What is the Department of Transportation shipping information for this product?
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How much Product G4510, Glutathione-Agarose, do I receive and how do I swell my powder?
1 answer-
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.
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How is Product G4510, Glutathione-Agarose, resin conjugated to the glutathione, and what is the binding capacity of the resin?
1 answer-
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.
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How do I regenerate Product G4510, Glutathione-Agarose, resin?
1 answer-
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.
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What reagents are compatible with Product G4510, Glutathione-Agarose, resin?
1 answer-
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.
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When using Product G4510, Glutathione-Agarose, how do I store the swollen resin?
1 answer-
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.
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