p27 Kip1 (Phospho-Ser10)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the measurement of p27 Kip1 phosphorylation at Ser10 in cell lysates. The kit targets p27 Kip1 (Phospho-Ser10) and is reactive with human, mouse, and rat samples, with a dynamic range of > 5000 Cells. Detection is achieved by a colorimetric method at 450 nm using an indirect ELISA format, in which the phospho-target is captured by specific primary antibodies and detected via HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition.

The assay is lysate-free and intended for adherent cell samples. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values, and the Crystal Violet whole-cell staining method may be applied after the colorimetric measurement to determine cell density. The kit can be used to measure the relative amounts of phosphorylated p27 Kip1 in cultured cells as well as to screen for the effects of various treatments, inhibitors, or activators on p27 Kip1 phosphorylation.

p27 Kip1, encoded by the gene CDKN1B, is a cell-cycle regulatory protein that interacts with cyclin-CDK2 and -CDK4, inhibiting cell cycle progression at G1 and acting as a tumour suppressor. It may also mediate TGF beta-induced G1 arrest, and its degradation is triggered by CDK-dependent phosphorylation and subsequent ubiquitination by SCF complexes. Phosphorylation of p27 Kip1 at Ser10 has been implicated in diseases including cancer, supporting applications in cancer biology, cell signalling, and drug discovery research.