OVCAR-3 human ovarian carcinoma cells, a HGSOC cell line [22 (link)], were provided by the RIKEN BioResource Research Center through the National BioResource Project of the Ministry of Education, Culture, Sports, Science and Technology /Japan Agency for Medical Research and Development, Japan. These cells were maintained in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% heat-inactivated fetal bovine serum (Biowest, Nuaillé, France), 100 U/mL penicillin, and 100 µg/mL streptomycin (Wako Pure Chemical, Osaka, Japan) at 37 °C in a 5% CO2 atmosphere. Anti-β-actin antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-ADP rabbit polyclonal antibody was obtained from LifeSpan Biosciences (Seattle, WA, USA). Rabbit polyclonal anti-phosphorylated Akt (S473) antibody, rabbit monoclonal anti-pan Akt antibody, and anti-pan extracellular signal-regulated kinase 1/2 (ERK1/2) antibody were obtained from Cell Signaling Technology (Beverly, MA, USA). Mouse monoclonal anti-di-phosphorylated ERK-1/2 was purchased from Sigma (St. Louis, MO, USA). The acyl-CoA-cholesterol acyltransferase 1 (ACAT-1) 1 inhibitor K604 and the hydroxymethylglutaryl-CoA reductase inhibitor pitavastatin were obtained from Sigma.
Anti di phosphorylated erk 1 2
Manufactured by Merck Group
Sourced in United States
Anti-di-phosphorylated ERK-1/2 is a laboratory reagent used to detect the dual phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) proteins. It is a specific antibody that binds to the activated, di-phosphorylated form of these kinases.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Penicillin, by Fujifilm (1 mentions)
Streptomycin, by Fujifilm (1 mentions)
Pitavastatin, by Merck Group (1 mentions)
Anti β actin antibody, by Santa Cruz Biotechnology (1 mentions)
C digit chemiluminescence scanner, by LI COR (1 mentions)
Anti total erk1, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti di phosphorylated erk 1 2
1
Ovarian Carcinoma Cell Culture Protocol
2
Western Blot Analysis of Phosphorylated ERK
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The samples were lysed in cold lysis buffer and the total homogenate (50 µg) was separated using 12% polyacrylamide gel. The proteins on nitrocellulose membrane were blocked with 5% non-fat dried milk and 0.1% Tween20 at RT and incubated overnight with primary antibodies 1/700 anti-diphosphorylated ERK1/2 (Sigma-Aldrich) and 1/1000 anti-total ERK1 (Santa Cruz Biotechnology, Inc). The blots were incubated with peroxidase-conjugated anti-rabbit (1/5000) or anti-mouse (1/2500 Jackson Immunoresearch Labs Inc) secondary antibodies and then revealed with ECL detection reagents (Inmun-Star HRP-Substrate Kits, Bio-Rad). Finally, the emitted light was captured by the C-DiGit Chemiluminescence Scanner (LI-COR Biosciences), and signals were quantified with ImageJ software.
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