Mouse VEGF ELISA Kit

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.

Target Information

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.

GENE ID 22339
SWISS PROT Q00731
SYNONYMS Vpf; Vegf

Materials Supplied

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

Storage Store at 2 - 8°C

Performance Characteristics

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