p53 (Phospho-Ser15)Colorimetric Cell-Based ELISA Kit

Assay Genie offers a Cell-Based ELISA Kit designed for the precise measurement of phosphorylated serine 15 on the p53 protein in cultured cells. The kit targets a key regulatory event on p53, a tumour suppressor protein, supporting research into p53 signalling pathways and the development of potential therapeutics.

Reactivity is established for human and rat samples, with a dynamic range of > 5000 Cells and a colorimetric detection method read at 450 nm. The kit is a lysate-free, high-throughput, sensitive assay that detects p53 protein phosphorylation and expression profile in cells, and can be used to measure relative amounts of phosphorylated p53 in cultured cells as well as to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on p53 phosphorylation. Qualitative determination of p53 (Phospho-Ser15) is achieved through an indirect ELISA format in which target-specific primary antibodies capture the analyte and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition.

Because the Cell-Based ELISA is qualitative, two normalisation methods are provided. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising target absorbance values, and following colorimetric measurement of HRP activity, Crystal Violet whole-cell staining may be used to determine cell density, allowing absorbance values to be normalised to cell amounts so plating differences can be adjusted.