Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for VEGF has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any VEGF present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for VEGF is added to the wells and binds to the combination of capture antibody-VEGF in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of VEGF present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven VEGF standard dilutions and VEGF sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Vascular endothelial growth factor (VEGF or VEGF-A), also known as vascular permeability factor (VPF), is a potent mediator of both angiogenesis and vasculogenesis in the fetus and adults. It is a member of the platelet derived growth factor family that is characterized by a cysteine knot structure formed by eight conserved cysteine residues. Mouse embryos expressing only the VEGF120 isoform do not survive to term and show defects in skeletogenesis. Mouse VEGF120 shares 98% aa sequence identity with corresponding regions of rat, 89% with canine, feline, equine and porcine, and 87% with human, ovine and bovine VEGF, respectively. VEGF binds the type I transmembrane receptor tyrosine kinases VEGF R1 (also called Flt 1) and VEGF R2 (Flk1/KDR) on endothelial cells. VEGF is required during embryogenesis to regulate the proliferation, migration, and survival of endothelial cells. In adults, VEGF functions mainly in wound healing and the female reproductive cycle. Pathologically, it is involved in tumor angiogenesis and vascular leakage. Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus. VEGF is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, oncostatin M and TNF-α.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse VEGF(8﹡12)||1 plate|
|Mouse VEGF Standard lyophilized, 2000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-mouse VEGF monoclonal antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|Standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 7pg/mL.|
|ASSAY RANGE||15.6-1000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant mouse VEGF. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant mouse||G-CSF, GM-CSF, IL-1α, IL-1γ, IL-2 , IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12, LIF, TNF-α, IFN-γ, MIP-2|
|Recombinant human||FIt-3 Ligand, PιGF, PDGF-AA, PDGF-AB|
Data Analysis Assistance
We have partnered with MyAssays to offer you an easy to use and versatile tool to analyze the data you receive using our ELISA Kit. Click the link below to be directed to the data analysis tool provided by MyAssays specifically for BSKM1018.