CD50/ICAM-3 (Phospho-Ser518)Colorimetric Cell-Based ELISA Kit

A Cell-Based ELISA Kit from Assay Genie designed for the qualitative detection of CD50/ICAM-3 phosphorylation at Ser518 in cultured cells. The kit is reactive with human samples and employs a colorimetric detection method at 450 nm, with a dynamic range of > 5000 Cells. It is formatted for high-throughput, lysate-free workflows and is intended for use with adherent cells.

Using an indirect ELISA format, the assay captures CD50/ICAM-3 (Phospho-Ser518) with target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling a colorimetric reaction upon substrate addition. The kit can be used to measure the relative amounts of phosphorylated CD50/ICAM-3 in cultured cells and to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on CD50/ICAM-3 phosphorylation.

A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values. After colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalized to cell amounts and correcting for plating differences. ICAM-3 is a cell surface glycoprotein of the immunoglobulin superfamily that plays a crucial role in immune response by facilitating leukocyte adhesion and activation, and phosphorylation at Ser518 has been implicated in regulating immune cell function and adhesion dynamics.