Western blot analysis was performed as previously described [19 (link)]. Briefly, MPBAs were treated with the indicated doses of Rg3 for two days. Total proteins were extracted using lysis buffer containing protease and phosphatase inhibitors, followed by processing for SDS-PAGE. The blots were incubated with the following primary antibodies as appropriate: PPARγ, perilipin, ERO1L, Phospho-eIF2α, CHOP, Phospho-Akt, total Akt, cleaved caspase3, total caspase3, and β-actin (all antibodies purchased from Cell Signaling Technology). An ECL Prime Western Blotting System (GE Healthcare) and ImageQuant LAS4000 (GE Healthcare) were used to detect protein bands. Densitometry analysis was performed using Image J software (NIH, Bethesda, MD, USA).
Ero1l
Manufactured by Cell Signaling Technology
Sourced in United States
ERO1L is an enzyme that catalyzes the oxidation of protein disulfide isomerase, which is involved in the formation of disulfide bonds in the endoplasmic reticulum. It plays a role in the regulation of the oxidative folding of proteins.
Automatically generated - may contain errors
Lab products found in correlation
Perilipin, by Cell Signaling Technology (1 mentions)
Ecl prime western blotting system, by GE Healthcare (1 mentions)
Phospho eif2α, by Cell Signaling Technology (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Total caspase 3, by Cell Signaling Technology (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence reagent kit, by Beyotime (1 mentions)
Na2so3, by Merck Group (1 mentions)
Caspase 9, by Proteintech (1 mentions)
Anti rabbit secondary antibody, by Proteintech (1 mentions)
Nahso3, by Merck Group (1 mentions)
Sds page gel preparation kit, by Beyotime (1 mentions)
Lats1, by Cell Signaling Technology (1 mentions)
Streptozotocin (stz), by MP Biomedicals (1 mentions)
Grp94, by Proteintech (1 mentions)
Eif2α, by Proteintech (1 mentions)
2 protocols using ero1l
1
Western Blot Analysis of Rg3 Signaling
2
Biochemical Assays for Oxidative Stress
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Na2SO3, NaHSO3 and HDX were purchased from Sigma-Aldrich (St. Louis, MO, USA). STZ was purchased from MP Biomedicals, LLC (Santa Ana, CA, USA). The antibodies for matrix metalloproteinase (MMP)9, MMP24, tissue inhibitor of metalloproteinase (TIMP)1 and GAPDH were purchased from Wuhan Boster Biological Technology, Ltd. (Wuhan, China). The dilution rate of these antibodies was 1:400. Furthermore, the antibodies for MST1, MST2, MOB1, LATS1, BIP, PDI and ERO1-L were purchased from Cell Signalling Technology (Danfoss, MA, USA). The antibodies for Bcl-2, Bax, caspase-3, caspase-9, CHOP, GRP94, eIF2α and GOT1 were purchased from Proteintech Group, Inc. (Chicago, IL, USA). The dilution rate of these antibodies was 1:1,000. Anti-rabbit secondary antibody and anti-rat secondary antibody were also purchased from Proteintech Group, Inc. The cell lysis buffer for western blot analysis, Bicinchoninic Acid (BCA) Protein Assay kit, Enhanced Chemiluminescence Reagent kit and SDS-PAGE Gel Preparation kit were obtained from Beyotime Institute of Biotechnology (Shanghai, China). The ELISA kit of SO2 was obtained from Mlbio Co. (Shanghai, China).
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