Human 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 well known for its important roles in regulating both physiological and pathological blood vessel growth. It is a member of the VEGF family that also includes VEGF-B, -C, -D, -E, and PlGF (placental growth factor). Family members exhibit a cysteine knot motif formed by eight characteristically spaced cysteine residues.

Various cells and tissues express VEGF including skeletal and cardiac muscle, hepatocytes, osteoblasts, neutrophils, macrophages, keratinocytes, platelets, brown adipose tissue, CD34+ cells, astrocytes, neurons, and endothelial cells. VEGF can be detected in both human plasma and serum samples, with higher levels found in serum due to its release from platelets. VEGF transcription is potentiated in response to hypoxia, oncogenic transformation, and growth factors.

VEGF has several physiological roles both during development and in the adult. It has a well-documented and critical role in embryonic vasculogenesis. Targeted deletion of a single VEGF allele results in significant defects in vascular development and is embryonic lethal. VEGF acts as an endothelial cell survival factor, mitogen, and inducer of migration, functions that potentially involve an array of signaling molecules including G proteins, PI3 kinase, MAP kinases, FAK, PKC, and NO. VEGF affects bone formation by regulating blood vessel growth and cartilage remodeling, activities important for growth plate morphogenesis. In the adult, VEGF has potential roles in cyclic angiogenesis in the female reproductive system. Other diverse functions include the regulation of hematopoietic development and roles as a neurotrophic factor.

VEGF has received much attention for its involvement in tumor-associated angiogenesis and its potential as a target for cancer therapy. Numerous studies have demonstrated that VEGF or its receptors are upregulated in several forms of human cancer. Neutralizing VEGF antibodies are shown to reduce tumor growth in cancer models.

GENE ID 7422
SWISS PROT P15692
SYNONYMS VPF; VEGF; MVCD1

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human VEGF (8﹡12) 1 plate
Human VEGF Standard lyophilized, 2000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-human 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 31.25 - 2000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant human VEGF. The factors listed below were prepared at 200ng/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 human EC-VEGF/PK1, PDGF-AA, PDGF-AB , PDGF-BB, PιGF, VEGF-B167, VEGF-C, VEGF-D, VEGF R3(Flt-4) /Fc Chimera
Recombinant mouse PιGF-2, VEGF120, VEGF164, VEGF-D, VEGF R2(KDR)/Fc Chimera, VEGF R3(Flt-4) /Fc Chimera
Other proteins Rat VEGF 120(0.07%), zebrafish VEGF 165