VEGFA ELISA Kit (Chicken)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Vascular Endothelial Growth Factor A (VEGFA) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Vascular Endothelial Growth Factor A (VEGFA) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin (By similarity) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Vascular Endothelial Growth Factor A (VEGFA) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Vascular Endothelial Growth Factor A (VEGFA) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Vascular Endothelial Growth Factor A (VEGFA), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Vascular Endothelial Growth Factor A (VEGFA) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 79-99 91 EDTA Plasma (n=5) 97-105 101 Heparin Plasma (n=5) 80-96 86 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 79-99 91 Serum (n=5) 79-99 91 EDTA Plasma (n=5) 97-105 101 EDTA Plasma (n=5) 97-105 101 Heparin Plasma (n=5) 80-96 86 Heparin Plasma (n=5) 80-96 86

Component Amount Anti-VEGFA Microplate 96 Wells (12 x 8 Well strips) VEGFA Lyophilized Standard 2 100X Biotinylated VEGFA Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-VEGFA Microplate 96 Wells (12 x 8 Well strips) Anti-VEGFA Microplate 96 Wells (12 x 8 Well strips) VEGFA Lyophilized Standard 2 VEGFA Lyophilized Standard 2 100X Biotinylated VEGFA Detector Antibody 1 x 120 uL 100X Biotinylated VEGFA Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Vascular Endothelial Growth Factor A (VEGFA) . No significant cross-reactivity or interference between Vascular Endothelial Growth Factor A (VEGFA) and analogues was observed.