Harvested cells were washed with Dulbecco’s phosphate-buffered saline (dPBS), centrifuged at 125 × g for 5 min and then the flash-frozen pellets were stored at −80 °C. Extracted RNA (500 ng–2 µg) were used as templates to make cDNA libraries using a High Capacity RNA-to-cDNA kit (Applied Biosystems). TaqMan gene expression assays were designed using GAPDH (Hs03929097, VIC) as an endogenous control and CDH1 (Hs01023895_m1, FAM), GSTP1 (Hs00943350_g1, FAM), HSPA1A (Hs00359163_s1, FAM), MGMT (Hs01037698_m1, FAM), MLH1 (Hs00179866_m1, FAM), PKP3 (Hs00170887_m1, FAM), RHOXF2 (Hs00261259_m1, FAM), SOX17 (Hs00751752_s1, FAM), SRPX2 (Hs00997580_m1, FAM), or THBS2 (Hs01568063_m1, FAM) as targets (Thermo Fisher Scientific). Quantitative PCR (qPCR) was performed in 10 µL reactions by using TaqMan Universal Master Mix II with UNG and 2 μL of diluted cDNA from each sample (Applied Biosystems). Gene expression levels were calculated by “delta delta Ct” and normalized to control samples using ViiA7 version 1.2.2 or QuantStudio version 1.3 software (Applied Biosystems by Life technologies).
Gstp1
Manufactured by Thermo Fisher Scientific
GSTP1 is a protein-based product designed for use in laboratory settings. It serves as a critical component in various biochemical and molecular biology applications. The core function of GSTP1 is to facilitate specific enzymatic reactions and interactions, though its precise intended use may vary depending on the specific research or experimental requirements.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh, by Thermo Fisher Scientific (1 mentions)
Taqman universal master mix 2 with ung, by Thermo Fisher Scientific (1 mentions)
Quantstudio version 1, by Thermo Fisher Scientific (1 mentions)
High capacity rna to cdna kit, by Thermo Fisher Scientific (1 mentions)
Thbs2, by Thermo Fisher Scientific (1 mentions)
Hspa1a, by Thermo Fisher Scientific (1 mentions)
Protein g sepharose bead, by GE Healthcare (1 mentions)
Protein g sepharose, by Thermo Fisher Scientific (1 mentions)
2 protocols using gstp1
1
Quantitative PCR Gene Expression Analysis
2
Immunoprecipitation Protocol for GSTP1
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Cells were lysed with NETN buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP-40, 1 mM EDTA, 1 % protease inhibitors cocktail). For the in vitro immunoprecipitation (IP) assay, the protein were purified for the assay. Briefly, the lysates or protein mixtures were precleaned by the protein G sepharose beads (GE Healthcare, USA) at 4 C for 1 hour. After that, the immunoprecipitation complex which containing protein lysates, blocked protein G sepharose beats and the corresponding antibodies (GSTP1, Thermo Fisher Scientific, Cat. # PA5-81507) or IgG, were incubated at 4 C overnight. All incubation processes needed to be done on a rotating shaker. The beads were collected and washed 4 times with the cold NETN buffer, which is ready for western blot assay after boiled in SDS loading buffer for 5 min.
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