p21 Cip1 (Phospho-Thr145)Colorimetric Cell-Based ELISA Kit

A high-throughput, lysate-free assay from Assay Genie designed for the quantitative detection of phosphorylated p21/Cip1 protein in cells. The kit uses a colorimetric detection method at 450 nm in an indirect ELISA format, in which p21 Cip1 (Phospho-Thr145) is captured by specific primary antibodies and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition. It is reactive with human, mouse, and rat samples and is intended for use with adherent cells.

The kit enables measurement of relative p21 Cip1 phosphorylation in cultured cells and screening of the effects of various treatments, inhibitors, or activators. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalizing target absorbance values, and Crystal Violet whole-cell staining is provided to determine cell density. The dynamic range is > 5000 Cells. The target is the CDKN1A gene product, a cyclin-dependent kinase inhibitor whose expression is tightly controlled by p53 and which mediates p53-dependent cell cycle arrest at G1.

Key components include 60 µL of 100x Anti-Phospho Target Antibody, 60 µL of 100x Anti-Target Antibody, 60 µL of Anti-GAPDH Antibody, 12 mL each of HRP-Conjugated Anti-Rabbit IgG Antibody and HRP-Conjugated Anti-Mouse IgG Antibody, 12 mL of Ready-to-Use Substrate, 50 mL of Blocking Buffer, 50 mL of 15X Wash Buffer, 24 mL of 10X TBS, 24 mL of Quenching Buffer, 24 mL of Stop Solution, 12 mL of SDS Solution, 12 mL of Crystal Violet Solution, 12 mL of Primary Antibody Diluent, and 2 adhesive plate seals.