Summary of FP assay for Akt
FP assay for Akt has been developed in 384-well plates using the same peptide substrate and anti-phosphoserine antibody as those used for PKC.
Concentration of the substrate is critical in development of FP assays due to specificity of the antibody.
Sensitivity of FP assay is ?-fold higher than that of SPA assay.
IC50’s of staurosporine determined in both FP and SPA assays are consistent.
Cost of reagents in the FP assay (10?per well in 384-well plates with a volume of 70 ?l) is less than that in the SPA assay.