CDC25B (Phospho-Ser323)Colorimetric Cell-Based ELISA Kit

This cell-based ELISA kit from Assay Genie is designed for the sensitive detection of phosphorylated CDC25B at the Ser323 residue. It uses an indirect ELISA format, in which CDC25B (Phospho-Ser323)-specific primary antibodies capture the target and HRP-conjugated secondary antibodies catalyze a colorimetric reaction measured at 450 nm. The assay is qualitative, intended for measuring relative levels of phosphorylated CDC25B in cultured cells and for screening the effects of treatments, inhibitors such as siRNA or chemicals, or activators on CDC25B phosphorylation.

A monoclonal anti-GAPDH antibody is included as an internal positive control for normalizing target absorbance, and a Crystal Violet whole-cell staining step allows cell density to be determined so that plating differences can be adjusted. The kit is validated for use with human, mouse, and rat samples and has a dynamic range of > 5000 cells, making it suitable for high-throughput work with adherent cells.

CDC25B is a member of the MPI phosphatase family that activates the cyclin-dependent kinase CDC2 by removing two phosphate groups and is required for entry into mitosis. Phosphorylation at Ser323 has been linked to its activation and to increased cell proliferation, making this phospho-specific ELISA a useful tool for research into cell cycle regulation, the G2/M transition, and potential therapeutic targets.