Recombinant Human GSTP1 Protein, CF Summary
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.|
|Shipping||The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- Assay Buffer: 100 mM NaH2PO4, pH 7.0
- Recombinant Human Glutathione S‑Transferase pi 1/GSTP1 (rhGSTP1) (Catalog # 6455-GT)
- Substrate: 1-bromo-2,4-dinitrobenzene (BDNB) (Sigma, Catalog # 262226), 75 mM stock in ethanol
- L-Glutathione, reduced (GSH) (Amresco, Catalog # 399), 250 mM stock in deionized water
- UV Plate (Costar, Catalog # 3635)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rhGSTP1 to 0.2 ng/μL in Assay Buffer.
- Dilute GSH to 4 mM in Assay Buffer.
- Combine equal volumes of 0.2 ng/μL rhGSTP1 and 4 mM GSH for 0.1 ng/μL rhGSTP1 with 2 mM GSH.
- Dilute Substrate to 2 mM in Assay Buffer.
- Load into a UV plate 50 μL of the rhGSTP1/GSH mixture. Include a substrate blank containing 25 μL of Assay Buffer with 25 μL of the 4 mM GSH prepared in step 2.
- Start the reaction by adding 50 μL of 2 mM Substrate to well.
- Read in kinetic mode for 5 minutes at an absorbance of 340 nm.
- Calculate specific activity:
Specific Activity (pmol/min/µg) =
|Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol|
|ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)|
*Adjusted for Substrate Blank
**Using the extinction coefficient 9600 M-1cm-1
***Using the path correction 0.320 cm
Note: the output of many spectrophotometers is in mOD. Per Well:
- rhGSTP1: 0.005 μg
- GSH: 1 mM
- Substrate: 1 mM
Background: Glutathione S-Transferase pi 1/GSTP1
Glutathione S-Transferases (GSTs) are members of the phase II detoxification enzyme family that conjugate glutathione to various electrophilic compounds, including metabolites generated by oxidative processes in the body, environmental toxins or carcinogens, and anti-cancer drugs. GSTP1 is a cytosolic protein that belongs to pi class of the GST superfamily. It is crystallized as a homodimer (1), but also exists in solution as an equilibrium mixture of monomer and dimer, depending on the protein concentration (2). Four genetic variants of GSTP1 with different enzymatic activities have been identified, which indicates the particular allelic form expressed in tissues could contribute to variation in catalytic efficiency and biological functions (3, 4). Human GSTP1 is present at elevated levels in many tumor cells, and has unique properties as a cancer marker (5). Genetic polymorphisms and expression patterns of GSTP1 have been associated with a variety of effects on human cancer, anti-cancer drug resistance, and asthma (6). In addition to its role as a drug-metabolizing enzyme, GSTP1 has ligand binding properties and regulates kinase signaling pathways through protein-protein interactions (7).
- Reinemer, P. et al. (1992) J. Mol. Biol. 227:214.
- Huang, Y.C. et al. (2008) J. Biol. Chem. 283:32880.
- Ali-Osman, F. et al. (1997) J. Biol. Chem. 272:10004.
- Hu, X. et al. (1998) Cancer Res. 58:5340.
- Sato, K. et al. (1992) Tohoku J. Exp. Med. 168:97.
- Townsend, D.M. and K.D. Tew (2003) Oncogene 22:7369.
- Adler, V. et al. (1999) EMBO J. 18:1321.
Product Specific NoticesCoomassie is a registered trademark of Imperial Chemical Industries Ltd.
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Reviews for Recombinant Human GSTP1 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human GSTP1 Protein, CF and earn rewards!
Have you used Recombinant Human GSTP1 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image