Background-subtracted data values (mean +/- SD) are graphed.
Untreated and staurosporine (ab120056) treated HeLa and Jurkat lysates were prepared in 1X Cell Extraction Buffer PTR and tested using the Cleaved PARP SimpleStepELISA. Raw OD 450 nm values are shown for 500 µg/mL lysate loads.
HeLa cells were treated with a dose titration of Staurosporine for 4 hours in complete media. Cells were cultured and treated in a 96-well cell culture microtiter plate. Lysates were prepared by direct in-well lysis without media removal: 2X Cell Extraction Buffer PTR was added to an equal volume of media and then resulting lysate was used directly in the Cleaved PARP SimpleStepTM ELISA assay. Raw values for triplicate measurements are plotted. The calculated IC50 is 0.77 µM.
20 µg of HeLa extracts that were untreated or treated for 4 hours with 1 µM Staurosporine were analyzed by western blot. The GAPDH blot is included to show the relative loads of each lysate. In the HeLa cell line, Staurosporine treatment is required to detect cleaved PARP protein, as observed in the SimpleStep ELISA (Figure 2).