Glutathione S-transferases (GSTs) are a family of enzymes that play an important role in detoxification by catalyzing the conjugation of many hydrophobic and electrophilic compounds with reduced glutathione. Based on their biochemical, immunologic, and structural properties, the soluble GSTs are categorized into 4 main classes: alpha, mu, pi, and theta. This GST family member is a polymorphic gene encoding active, functionally different GSTP1 variant proteins that are thought to function in xenobiotic metabolism and play a role in susceptibility to cancer, and other diseases. In response to oxidative stress, upregulation of the GST family member GSTP1 occurs, consistent with this function. Furthermore, the GSTP1 gene is subject to CpG island hypermethylation, a state that correlates with human prostatic carcinogenesis. GSTP1 gene hypermethylation can be detected in urine, ejaculate and plasma from men with prostate cancer, potentially making GSTP1 a useful biomarker for prostate cancer screening.
Background References
1. Sun K H et al. Glutathione-S-transferase P1 is a critical regulator of Cdk5 kinase activity. J Neurochem 118:902-914 (2011).
2. Kong K H et al. Tyrosine-7 in human class Pi glutathione S-transferase is important for lowering the pKa of the thiol group of glutathione in the enzyme-glutathione complex. Biochem Biophys Res Commun 184:194-197 (1992).
Western blot analysis of GST3 on different lysates with Mouse anti-GST3 antibody (EM1801-18) at 1/2,000 dilution.
Lane 1: HeLa cell lysate Lane 2: HEK-293 cell lysate Lane 3: MDA-MB-231 cell lysate Lane 4: HT-29 cell lysate Lane 5: Jurkat cell lysate Lane 6: K-562 cell lysate Lane 7: JAR cell lysate Lane 8: Human brain tissue lysate Lane 9: Human lung tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 23 kDa Observed band size: 23 kDa
Exposure time: 24 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1801-18) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP, AlpSdAbs® VHH(HRP) (001-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-GST3 antibody (EM1801-18) at 1/10,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1801-18) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
☑ Knockdown (KD)
Western blot analysis of GST3 on different lysates with Mouse anti-GST3 antibody (EM1801-18) at 1/1,000 dilution.
Lane 1: HCT 116-si NT cell lysate Lane 2: HCT 116-si GST3 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 23 kDa Observed band size: 23 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1801-18) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
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