The cells were homogenized in lysis buffer (50 mM/L HEPES pH 7.5, 150 mM/L NaCl, 10% glycerol, 1% Triton X-100, 1 mM/L EGTA, 1.5 mM/L MgCl2, 10 mM/L NaF, 10 mM/L sodium pyrophosphate, 1mM/L Na3VO4, 10 μg approtinin/ml, 10 μg leupeptin/ml) (Sigma). The following antibodies were used: α-RAS, α-RAC, α-CDC42, α-RHO, α-pEGFR, α-EGFR, α-pFAK, α-FAK, α-pSCR, α-SCR, α-pMEK, α-MEK, α-pERK1/2, α-ERK1/2, α-pAKT, α-AKT, α-pGSK3 α /β, α-GSK3 α /β, α-β catenin, α-pCHK2, α-CHK2, α-tubulin, α-SP1, α-pRET, α-RET (Cell Signaling, Danvers, MA, USA), and α-SOD3 (Santa Cruz, Santa Cruz, CA, USA). Signal density analysis was performed using ImageJ software GEL blot software.
α mek
Manufactured by Cell Signaling Technology
Sourced in United States
α-MEK is a primary antibody that specifically recognizes mitogen-activated protein kinase kinase (MEK), a key component of the MAPK signaling pathway. This antibody can be used for the detection and analysis of MEK in various applications such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
α erk1 2, by Cell Signaling Technology (2 mentions)
α pchk2, by Cell Signaling Technology (1 mentions)
α akt, by Cell Signaling Technology (1 mentions)
α egfr, by Cell Signaling Technology (1 mentions)
α fak, by Cell Signaling Technology (1 mentions)
α perk1 2, by Cell Signaling Technology (1 mentions)
α β catenin, by Cell Signaling Technology (1 mentions)
α sp1, by Cell Signaling Technology (1 mentions)
α tubulin, by Cell Signaling Technology (1 mentions)
Leupeptin, by Merck Group (1 mentions)
α flag m2, by Merck Group (1 mentions)
α phospho erk1 2, by Cell Signaling Technology (1 mentions)
α gapdh, by Merck Group (1 mentions)
Ecl plus, by GE Healthcare (1 mentions)
Tween 20, by Merck Group (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
2 protocols using α mek
1
Western Blot Analysis of Cell Signaling
2
Western Blot Analysis of Phospho-Proteins
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Cleared lysates were adjusted to equal protein concentrations after protein quantification using the Bradford assay (Biorad), separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretically transferred to PVDF membranes. Membranes were blocked in 5% non-fat milk in TBS containing 0.1% Tween20 (Sigma-Aldrich) and incubated with the following primary antibodies overnight: The rabbit polyclonal α-phospho-S614 antibody which was produced by Eurogentec against the phosphorylated version of the HQFEQLS(P)GSILW peptide and affinity purified against this peptide, α-FLAG M2 (Sigma Aldrich), α-phospho-Erk1/2, α-Erk1/2, αphospho-MEK, α-MEK (Cell Signalling), α-GAPDH (Merck Millipore), α-14-3-3ε and α-CRAF C-12 (Santa Cruz Biotechnology). Membranes were washed and then incubated one hour at ambient temperature with species-macthed secondary antibodies (Jackson Immunoresearch). Protein bands were visualized by ECL plus (GE Healthcare).
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