VEGFA : VEGFR2 [Biotinylated] Inhibitor Screening ELISA Kit

背景（Background）

Kinase insert domain receptor (KDR) is also known as CD309, FLK1, VEGFR, VEGFR2, and is one of the subtypes of VEGFR. VEGF receptors are receptors for vascular endothelial growth factor (VEGF). There are three main subtypes of VEGFR, numbered 1, 2 and 3. The VEGF receptors have an extracellular portion consisting of 7 immunoglobulin-like domains, a single transmembrane spanning region and an intracellular portion containing a split tyrosine-kinase domain. VEGF-A binds to VEGFR-1 (Flt-1) and VEGFR-2 (KDR/Flk-1). VEGFR-2 appears to mediate almost all of the known cellular responses to VEGF.The function of VEGFR-1 is less well defined, although it is thought to modulate VEGFR-2 signaling. Another function of VEGFR-1 may be to act as a dummy/decoy receptor, sequestering VEGF from VEGFR-2 binding (this appears to be particularly important during vasculogenesis in the embryo). In addition, VEGFR2 is able to interact with HIV-1 extracellular Tat protein upon VEGF activation, and seems to enhance angiogenesis in Kaposi's sarcoma lesions.

应用说明（Application）

This kit is developed for screening for inhibitors of human VEGFA binding to human VEGFR2.

It is for research use only.

重构方法（Reconstitution）

Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.

存储（Storage）

Unopened kit should be stored at 2°C-8°C upon receiving. Find the expiration date on the outside packaging and do not use reagents past their expiration date.

The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

原理（Assay Principles）

This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new VEGFA pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human VEGFR2 to immobilized human VEGFA in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated VEGFR2 protein, a human VEGFA protein, an anti-VEGFA neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.

Your experiment will include 4 simple steps:

a) Coat the plate with human VEGFA.

b) Add your molecule of interest to the tests.

c) Add human VEGFR2-Biotin to bind the coated VEGFA.

d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.

Finally, the half maximal inhibitory concentration (IC50) of your compound to VEGFA: VEGFR2 binding will be determined by comparing OD readings among different experimental groups.