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Recombinant Human GSTM1 Protein, CF

Catalog #: 6894-GT Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
6894-GT-010
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human GSTM1 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 >20,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Glutathione S-Transferase mu 1/GSTM1 protein
Met1-Lys218
Accession #
P09488
N-terminal Sequence
Analysis
Met1 and Pro2
Predicted Molecular Mass
26 kDa
SDS-PAGE
25-27 kDa, reducing conditions

Product Datasheets

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6894-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.

6894-GT

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT 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: 50 mM Sodium Phosphate, pH 7.0
  • Recombinant Human Glutathione S‑Transferase mu 1/GS (rhGSTM1) (Catalog # 6894-GT)
  • L-Glutathione, reduced (GSH) (Amresco, Catalog # 0399), 250 mM stock in deionized water
  • Substrate: 1-Bromo-2,4-dinitrobenzene (BDNB) (Sigma, Catalog # 262226), 75 mM stock in ethanol
  • UV plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute GSH to 1 mM in Assay Buffer.
  2. Dilute rhGSTM1 to 0.2 ng/μL in Assay Buffer containing 1 mM GSH.
  3. Dilute Substrate to 2 mM in Assay Buffer containing 1 mM GSH.
  4. Load into a plate 50 μL of 0.2 ng/μL rhGSTM1 and start the reaction by adding 50 μL of 2 mM Substrate. For Substrate Blanks, load 50 μL of Assay Buffer containing 1 mM GSH and 50 μL of 2 mM Substrate.
  5. Read plate at a wavelength of 340 nm (bottom read) in kinetic mode for 5 minutes.
  6. 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 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhGSTM1: 0.010 μg
  • Substrate: 1 mM
  • GSH: 1 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Glutathione S-Transferase mu 1/GSTM1

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. GSTM1 is a cytosolic protein that belongs to the mu class of the GST superfamily. The gene encoding GSTM1 is mapped onto human chromosome 1p13.3 and is known to be highly polymorphic (1). Mostly notably, the widely occurring GSTM1-null genotype has been linked to a variety of cancers including lung (2), gastric (3), bladder (4) and prostate (5). In addition to its role in releasing oxidative stress, GSTM1 has also been suggested to act as a hormone binding protein and play a role in maintaining hormone homeostasis in the body (6, 7).

References
  1. Pearson, W. R. et al. (1993) Am. J. Hum. Genet. 53:220.
  2. Mohr, L.C. et al. (2003) Anticancer. Res. 23:2111.
  3. Wang, H. et al. (2010) Dig. Dis. Sci. 55:1824.
  4. Engel, L. S. et al. (2002) Am. J. Epidermiol. 156:95.
  5. Mo, Z. et al. (2009) Prostate. 69:662.
  6. Mukherjee, S. B. et al. (1999) Biochem. J. 340:309.
  7. Ishigaki, S. et al. (1989) Arch. Biochem. Biophys. 273:265.
Entrez Gene IDs
2944 (Human)
Alternate Names
EC 2.5.1.18; glutathione S-alkyltransferase; glutathione S-aralkyltransferase; glutathione S-aryltransferase; glutathione S-transferase M1; Glutathione STransferase mu 1; Glutathione S-Transferase mu 1; GST class-mu 1; GST HB subunit 4; GST1; GSTM1; GSTM1-1; GSTM1a-1a; GSTM1b-1b; GTH4; GTM1; HB subunit 4; H-B; MGC26563; MU; MU-1; S-(hydroxyalkyl)glutathione lyase

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