IRAK1 (Phospho-Thr209)Fluorometric Cell-Based ELISA Kit

This Fluorometric Cell-Based ELISA Kit from Assay Genie is a lysate-free, high-throughput and sensitive assay for monitoring IRAK1 phosphorylation and expression profile in cells. The kit can be used to measure the relative amounts of phosphorylated IRAK1 (Phospho-Thr209) in cultured cells, and to screen the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on IRAK1 phosphorylation.

Detection is achieved by an indirect ELISA format in which IRAK1 (Phospho-Thr209) is captured by phospho-specific primary antibodies, while Dye 1- and Dye 2-conjugated secondary antibodies bind the Fc region of the primary antibody to enable a fluorometric readout. The Dynamic Range is > 5000 Cells. The kit is reactive with Human samples, with storage and stability specified as 4°C/6 Months. The format comprises two 96-well plates. A fluorescent plate reader with two channels at Ex/Em 651/667 and 495/521 is required but not provided. Database identifiers for the target include Gene ID 3654, UniProt P51617, OMIM 300283 and Unigene Hs.200716/Hs.522819.

Two normalization strategies are supported: a monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target RFU values, and an antibody against the non-phosphorylated counterpart of IRAK1 (Phospho-Thr209) is provided so that RFU values for non-phosphorylated IRAK1 can be used to normalize the RFU value for phosphorylated IRAK1. IRAK1 is a serine/threonine kinase of the IRAK family involved in Toll/IL-1 signaling, associating with the interleukin-1 receptor upon IL-1 engagement to trigger downstream cascades, and is partially responsible for IL-1-induced upregulation of the transcription factor NF-kappa B. The human gene maps to chromosomal location Xq28.