Lck (Phospho-Tyr505)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie enables qualitative detection of Lck phosphorylated at tyrosine 505 directly in cultured cells, without the need for lysate preparation. The assay uses an indirect ELISA format in which phospho-specific primary antibodies capture the target, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyze a colorimetric reaction upon substrate addition. Detection is colorimetric at 450 nm.

The kit has a stated dynamic range of > 5000 Cells and shows reactivity with Human, Mouse and Rat samples. It is suitable for adherent cells and supports high-throughput work, allowing researchers to measure relative amounts of phosphorylated Lck in cultured cells or to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on Lck phosphorylation. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values, and following the colorimetric readout the Crystal Violet whole-cell staining method may be used to determine cell density, with absorbance values normalized to cell amounts to adjust for plating differences.

Phosphorylation of Lck (Lymphocyte-specific protein tyrosine kinase) at tyrosine 505 is known to influence Lck activity and downstream signaling, including phosphorylation of ITAM tyrosines within the cytoplasmic tails of TCR gamma chains and CD3 subunits, making this phospho-specific kit a relevant tool for research into T-cell receptor signaling, immune regulation, autoimmune disease and cancer.