p27 Kip1 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA is designed for the quantitative measurement of p27 Kip1 levels in cell lysates and tissue homogenates, using a colorimetric detection method at 450 nm. The kit targets human p27 Kip1 (gene symbol CDKN1B) and is supplied in a 96-Well Microplate format intended for use with adherent cells. It has a dynamic range of > 5000 Cells and provides a convenient, lysate-free, high-throughput approach to detecting the p27 Kip1 protein expression profile in cultured cells.

The assay uses an indirect ELISA format: p27 Kip1 is captured by p27 Kip1-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. On substrate addition, the conjugated HRP enzyme catalyses a colorimetric reaction. Researchers can use the kit to measure relative amounts of p27 Kip1 in cultured cells and to assess the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on p27 Kip1.

Because the Cell-Based ELISA is qualitative, two normalisation methods are provided. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used to determine cell density and adjust for plating differences. The kit is stored at 4 °C with a stated stability of 6 Months.