Cells were harvested in RIPA buffer (Thermo Scientific) with protease and phosphatase inhibitors (Roche Diagnostics). Immunoblotting was done after sodium dodecyl sulfate–polyacrylamide gel electrophoresis on gradient 4–15% gels (Biorad) at 30 μg protein and transferred to polyvinylidene fluoride membranes. All antibodies besides the secondary horseradish peroxidase-conjugated antibody (Santa Cruz Biotechnology) were purchased from Cell Signaling Technologies: phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204), p44/42 MAPK (ERK1/2), phospho-p38 (Thr180/Tyr182), p38, phospho- MEK (Ser 217/221), MEK, RhoC, RhoA, ß-actin. SuperSignal West Pico Luminol/Enhancer Solution was purchased from Thermo Scientific.
Phospho mek ser 217 221
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-MEK (Ser 217/221) is a primary antibody that specifically recognizes MEK1 and MEK2 when phosphorylated at Ser217 and Ser221. This product is suitable for Western blotting applications.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Supersignal west pico luminol enhancer solution, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
Sapanisertib, by Selleck Chemicals (1 mentions)
Total mek, by Cell Signaling Technology (1 mentions)
Trametinib, by Selleck Chemicals (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Phospho p70 s6 kinase thr389, by Cell Signaling Technology (1 mentions)
Nf1 a300 140a, by Fortis Life Sciences (1 mentions)
Phospho rps6 ser235 236, by Cell Signaling Technology (1 mentions)
Phospho 4e bp1 thr37 46, by Cell Signaling Technology (1 mentions)
Phospho akt thr308, by Cell Signaling Technology (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Phospho erk thr202 tyr204, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 protocols using phospho mek ser 217 221
1
Immunoblotting for MAPK Signaling
2
Quantification of Signaling Proteins
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Antibodies against total ERK, phospho-ERKthr202/tyr204, total MEK, phospho-MEKser217/221, phospho-p70-S6 kinasethr389, phospho-RPS6ser235/236, cleaved PARP, B-actin, phospho-4EBP1thr37/46, total AKT, phospho-AKTser473, and phospho-AKTthr308 were obtained from Cell Signaling Technology, NF1 (A300-140A) from Bethyl, and Cyclin D1 from Santa Cruz Biotechnology. Trametinib and sapanisertib were purchased from SelleckChem. Drugs for in vitro studies were dissolved in DMSO to yield 10 mM or 1 mM stock solutions and stored at −20°C.
3
Antibody-based Signaling Pathway Analysis
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RPMI-1640 Medium, Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco (Grand Island, NY, USA). Antibodies against phospho-ERK (Thr202/Tyr204; #4370, 1:1,000), ERK (#4695, 1:1,000 WB), phospho-MEK (Ser217/221; #9154, 1:2,000), MEK (#9122, 1:2,000), were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against tubulin (#66240-1-Ig, 1:5,000), β-actin (#60008-1-Ig, 1:5,000), were purchased from Proteintech. The NCOA2 (ab1877, 1:5,000) antibody was purchased from Abcam (Cambridge, UK). Secondary antibodies (1:5,000) conjugated to horseradish peroxidase (HRP) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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