CrkL (Phospho-Tyr207)Fluorometric Cell-Based ELISA Kit

A Fluorometric Cell-Based ELISA Kit from Assay Genie is designed to monitor the phosphorylation and expression profile of the CRKL protein at tyrosine 207 in cultured cells. Described as a lysate-free, high-throughput and sensitive assay, it allows qualitative determination of the target through an indirect ELISA format in which target-specific primary antibodies are detected by Dye 1- and Dye 2-conjugated secondary antibodies that emit light upon excitation, enabling a fluorometric readout. The assay is suitable for measuring the relative amounts of phosphorylated CrkL in cultured cells and for screening the effects of various treatments, inhibitors such as siRNA or chemicals, and activators on CrkL phosphorylation.

Two normalization strategies are built into the workflow: a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalizing target RFU values, and an antibody against the non-phosphorylated counterpart of the target is provided so that the RFU values for non-phosphorylated CrkL can be used to normalize the RFU value for phosphorylated CrkL. The kit is reactive with human, mouse and rat samples and is read on a fluorescent plate reader at Ex/Em of 651/667 (Dye 1) and 495/521 (Dye 2). The stated dynamic range is > 5000 Cells, storage is 4°C with a stability of 6 Months, and database cross-references include Gene ID 1399, UniProt ID P46109, OMIM 602007 and Unigene Hs.592205.