CDC2 (Phospho-Tyr15)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is specifically designed for the detection of phosphorylated CDC2 (CDK1) at tyrosine 15 in cell lysates, using adherent cells as the sample type. The assay uses a colorimetric detection method at 450 nm and is reactive with human, mouse, and rat samples, with a dynamic range of > 5000 cells. CDC2 is a protein kinase of the CDK family that serves as the catalytic subunit of the M-phase promoting factor (MPF), which is essential for G1/S and G2/M phase transitions. Phosphorylation of CDC2 at tyrosine 15 regulates its activity and function, making it a critical marker for studying cell cycle regulation and cellular proliferation. The gene encoding this target is activated in many cancers including colon, liver, and breast.

The kit employs a lysate-free, high throughput, and sensitive indirect ELISA format, in which target-specific primary antibodies capture the phosphorylated CDC2, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Through this binding, the HRP enzyme conjugated to the secondary antibody can catalyze a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target absorbance values. In addition, following the colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method may be used to determine cell density, and the absorbance values can be normalised to cell amounts to adjust for plating differences.

Researchers can use this kit to quantify phosphorylated CDC2 levels in cultured cells, as well as to screen for the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on CDC2 phosphorylation. Database information from the manufacturer lists Gene ID: 983, UniProt ID: P06493, OMIM: 116940, Unigene: Hs.334562, with the gene symbol CDK1 and sub type Phospho.