VEGFR1 (Phospho-Tyr1333)Fluorometric Cell-Based ELISA Kit

An Assay Genie cell-based phospho ELISA designed to monitor VEGFR1 (Phospho-Tyr1333) protein phosphorylation and expression profile in cultured cells. The kit is convenient, lysate-free and high-throughput, allowing measurement of relative amounts of phosphorylated VEGFR1 in cells, and screening for the effects of various treatments, inhibitors such as siRNA or chemicals, or activators on VEGFR1 phosphorylation (FLT1).

Qualitative determination is achieved by an indirect ELISA format in which VEGFR1 (Phospho-Tyr1333) is captured by target-specific primary antibodies, while Dye 1-conjugated and Dye 2-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling fluorometric detection. Two normalisation reagents are included: a monoclonal antibody specific for human GAPDH as an internal positive control, and an antibody against the non-phosphorylated counterpart of VEGFR1 (Phospho-Tyr1333) for normalising RFU values. The assay is reactive with human, mouse and rat samples, with a dynamic range of > 5000 Cells, and is stored at 4°C for 6 Months.

The kit provides clear-bottom black plates together with reagents for fixation, quenching, blocking, washing and detection. Recommended fluorescent plate reader channels are Ex/Em 651/667 (Dye 1) and 495/521 (Dye 2). The target gene is FLT1 (Gene ID: 2321; UniProt: P17948), a receptor tyrosine kinase of the VEGFR family that binds VEGF, VEGFB and PGF and plays a key role in vascular development and regulation of vascular permeability.