This antibody is specific for human RSK1 p90 phoshorylated on Threonine 359 and Serine 363.
80% identities with RSK3 p90 (pT365+pS369), 75% with RSK2 p90 (pT356+pS360) and 74% identities with RSK6 p90 (pT368+pS372)
Western blot - Anti-RSK1 p90 (phospho T359 + S363) antibody [E238] (ab32413)
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 90 kDa
Exposure time : 16 minutes
BT549 cells were incubated at 37 °C for 1h with vehicle control (0 μM) and different concentrations of PD 173074 (ab141117) in DMSO. Decreased expression of RSK1 p90 (phospho T359 + S363) (ab32413) correlates with an increase in PD 173074 concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab32413 at 1/5000 dilutionand ab8227 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
Western blot - RSK1 p90 (phospho T359 + S363) antibody [E238] (ab32413)
All lanes : Anti-RSK1 p90 (phospho T359 + S363) antibody [E238] (ab32413) at 1/20000 dilution
Lane 1 : A431 cell lysate untreated. Lane 2 : A431 cell lysate treated with EGF.
Predicted band size : 90 kDa Observed band size : 90 kDa