Western Blot and Kinase Assays

Western blot, immunoprecipitation, and in vitro kinase assays were performed as previously described (11 (link)). The following antibodies were used: anti-p217/p221-MEK1/2 (p-MEK1/2) (#9154), anti-p202/p204-ERK1/2 (p-ERK1/2) (#4370), anti-MEK1/2 (#4694), anti-ERK1/2 (#4696), anti-p380-p90RSK(p-RSK) (#9341), GAPDH (#2118) from Cell Signaling, anti-V5 (R960–25) from Thermo Fisher Scientific and anti-BRAF (sc-5284) and anti-cyclin D1 (M-20) from Santa Cruz Biotechnology.

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Gao Y., Maria A., Na N., Paula A.D., Gorelick A.N., Hechtman J.F., Carson J., Lefkowitz R.A., Weigelt B., Taylor B.S., Zhao H., Reis-Filho J.S., de Stanchina E., Rosen N., Yao Z, & Yaeger R. (2019). V211D mutation in MEK1 causes resistance to MEK inhibitors in colon cancer. Cancer discovery, 9(9), 1182-1191.

Publication 2019

anti-p217/p221-MEK1/2 (p-MEK1/2) anti-p202/p204-ERK1/2 (p-ERK1/2) anti-MEK1/2 anti-ERK1/2 GAPDH anti-V5 (R960–25) anti-BRAF (sc-5284) anti-cyclin D1 (M-20)

Antibodies Assays Braf Cyclin d1 Erk1 Gapdh Immunoprecipitation Kinase Mek1 P90rsk Western blot

Corresponding Organization : Memorial Sloan Kettering Cancer Center

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Variable analysis

independent variables

Western blot, immunoprecipitation, and in vitro kinase assays

dependent variables

Levels of p217/p221-MEK1/2 (p-MEK1/2)
Levels of p202/p204-ERK1/2 (p-ERK1/2)
Levels of MEK1/2
Levels of ERK1/2
Levels of p380-p90RSK(p-RSK)
Levels of GAPDH
Levels of V5
Levels of BRAF
Levels of cyclin D1

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

Annotations

Based on most similar protocols

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