Product Overview
ID Components Size (96 tests)
ISK-022-1 High-bind Plate 1 plate
ISK-022-2 Human VEGFA 10 μg
ISK-022-3 Biotinylated Human VEGFR2 20 μg
ISK-022-4 Anti-VEGFA Neutralizing Antibody 20 μg
ISK-022-5 Streptavidin-HRP 10 μg
ISK-022-6 Coating Buffer 12 mL
ISK-022-7 10xWashing Buffer 50 mL
ISK-022-8 Blocking Buffer 50 mL
ISK-022-9 Substrate Solution 12 mL
ISK-022-10 Stop Solution 7 mL
Description
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 Kaposis sarcoma lesions.
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.