Recombinant Human GSTP1 Protein, CF

Catalog #: 6455-GT Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
6455-GT-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human GSTP1 Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

Measured by the conjugation of reduced glutathione to 1-bromo-2,4-dinitrobenzene. The specific activity is >25,000 pmol/min/μg, as measured under the described conditions.

Source

E. coli-derived human Glutathione S-Transferase pi 1/GSTP1 protein
Met1-Glu210

Accession #

AAC51280

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

23 kDa

SDS-PAGE

24 kDa, reducing conditions

Product Datasheets

6455-GT

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

6455-GT

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. 6 months from date of receipt, -70 °C as supplied. 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials

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 V_max* (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

Reconstitution Calculator

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).

References

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.

Entrez Gene IDs

2950 (Human)

Alternate Names

deafness, X-linked 7; DFN7; EC 2.5.1.18; FAEES3; fatty acid ethyl ester synthase III; glutathione S-transferase P; Glutathione STransferase pi 1; Glutathione S-Transferase pi 1; GST class-pi; GST3DFN7; GSTP; GSTP1; GSTP1-1; PI