ISK-025

VEGFA; KDR

ID Components Size (96 tests)
ISK-025-1 High-bind Plate 1 plate
ISK-025-2 Human VEGF R2 50 μg
ISK-025-3 Biotinylated Human VEGFA 10 μg
ISK-025-4 Anti-VEGF Neutralizing Antibody 20 μg
ISK-025-5 Streptavidin-HRP 10 μg
ISK-025-6 Coating Buffer 12 mL
ISK-025-7 10xWashing Buffer 50 mL
ISK-025-8 Blocking Buffer 50 mL
ISK-025-9 Substrate Solution 24 mL
ISK-025-10 Stop Solution 7 mL

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.

96 tests

20-0.01μg/mL

This inhibitor screening ELISA kit is designed to facilitate the identification and characterization of new VEGF165 pathway inhibitors. The assay takes advantage of our in house-developed binding of biotinylated human VEGF165 to immobilized human VEGF R2 in a functional ELISA assay, and employs a simple colorimetric ELISA platform. Briefly, we provide you with a human Biotinylated VEGF165 protein, a human VEGF R2 protein, an anti-VEGF neutralizing antibody (as method verified Std.), and Streptavidin-HRP reagent.
Your experiment will include 4 simple steps:
a) Coat the plate with human VEGF R2.
b) Add your molecule of interest to the tests.
c) Add human VEGF165-Biotin to bind the coated VEGF R2.
d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
Finally, the half maximal inhibitory concentration (IC50) of your compound to VEGF165: VEGF R2 binding will be determined by comparing OD readings among different experimental groups.

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

See Certificate of Analysis for details of reconstitution instruction and specific concentration.

The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.

Serial dilutions of Anti-VEGFA Neutralizing antibody (1:1 serial dilution, from 20 μg/mL to 0.01μg/mL) was added into Biotinylated VEGFA: VEGF R2 binding reactions. The assay was performed according to the protocol described below. Background was subtracted from data points prior to log transformation and curve fitting.