As shown, using the p38 MAPKalpha assay kits, a significant stimulation of p38 MAPKalpha phosphorylation at Thr180/Tyr182 is detected in HEK293 cells treated with anisomycin for 30 minutes compared with untreated cells, while no change in total p38 MAPKalpha levels is observed.
Using the PhosphoTracer phospho-p38 MAPKalpha assay, the length of assay incubation time that was required to detect recombinant phospho-p38 MAPKalpha was examined. After 15 mins, phospho-p38 MAPKalpha concentrations of approximately 4 ng/mL were readily detected, while after 1 hour incubation, the limit of detection was approximately 1 ng/mL, and with a much improved assay window. Incubation times longer than one hour showed a slight improvement on assay performance.
PhosphoTracer p38 MAPKalpha (pT180/Y182) + total p38 MAPKalpha ELISA Kit (ab119665) used in Sandwich ELISA.