As shown, using the GAPDH assay kit, cellular GAPDH is readily detected in A431 cellular lysates, in either untreated cells (-), or cells treated with EGF (+).
Various concentrations of recombinant GAPDH diluted in Lysis Mix containing 0.1% BSA were analyzed using the PhosphoTracer cellular GAPDH assay. The recombinant GAPDH dilutions were transfered to an PhosphoTracer assay plate and analyzed using the standard PhosphoTracer assay protocol. Signal in the wells was determined using a plate reader.
Using the PhosphoTracer GAPDH assay, the amount of lysate required to detect GAPDH in HeLa cell lysates was examined. Up to 50 µg of lysate/well was loaded into replicate wells of an PhosphoTracer microplate, and analyzed for GAPDH. GAPDH was readily detected in less than 1 µg of lysate/well.