VEGF ELISA Kit, Human

Catalog number: KHG0112

Novex™  Related applications: Protein Assays and Analysis

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Description

The Human Vascular Endothelial Growth Factor (VEGF) ELISA research-use-only kit is to be used for the quantitative determination of VEGF in samples (see sample types indicated) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant forms of this target.

Performance characteristics

• Sensitivity: <5 pg/mL
• Standard curve range: 23.4‒1,500 pg/mL
• Sample type(s): serum, plasma, cell culture supernatant
• Specificity: natural and recombinant human VEGF
• Cross-reactivity: see kit manual for cross-species and/or cross-target reactivity
• Sample volume: 50 μL
• Total assay time: 4 hours

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Principle of the method

The human VEGF solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Target information

Vascular Endothelial Growth Factor (VEGF), originally named vascular permeability factor (VPF), is an important regulator of angiogenesis and vasculogenesis. Angiogenesis occurs in normal processes related to the female reproductive cycle and in pathological processes such as tumor growth and metastasis, diabetic retinopathy, rheumatoid arthritis, or after tissue ischemia. Vasculogenesis involves the formation of blood vessels through the differentiation of endothelial cells from mesodermal precursors. Whereas vasculogenesis is restricted to embryonic development, angiogenesis operates throughout life when new vascularization is required.

Alternative splicing of VEGF mRNA accounts for 5 isoforms of 121, 145, 165, 189, and 206 amino acids, of which the 165 amino acid form is the most abundant. The active forms of these isoforms are disulfide-linked homodimers. VEGF-121 differs from the larger VEGF isoforms in that it is the only one that does not bind to heparin. The 165 amino acid protein is a 46 kDa homodimer, which is produced by numerous cell types that include a variety of tumors, macrophages and folliculostellate cells.

There are two known high-affinity receptors for VEGF, fms-like tyrosine kinase (Flt-1) and fetal liver kinase (Flk-1). Both are tyrosine kinases and appear to be exclusively expressed on endothelial cells. The binding of VEGF to its receptor activates a signaling cascade that results in the activation of the mitogen-activated kinase (MAPK) and the tyrosine phosphorylation of phospholipase Cγ1 (PLCγ 1), which in turn leads to increases in the intracellular levels of inositol 1,4,5-triphosphate and calcium. The increase in calcium triggers nitric oxide synthase (NOS) to produce NO. This NOS activity is necessary for VEGF to stimulate angiogenesis and increase vascular permeability. Expression of VEGF and its receptor occurs during angiogenesis and vasculogenesis.

Related links

Learn more about ELISA kits
Learn more about other immunoassays

For Research Use Only. Not for use in diagnostic procedures.

Specifications

Gene Aliases: MGC70609, MVCD1, VEGF, VPF
Shipping Condition: Wet Ice
Marker: VEGF
Gene ID: 7422
Species: Human
Gene Symbol: VEGFA
Sensitivity: <5 pg⁄mL
Target Gene: vascular endothelial growth factor A
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 192 assays
Concentration: 23.4-1500 pg⁄ml
Sample Volume: 50 µl
Protein Family: Growth Factors & Receptors
Incubation Time: 4 hrs
Detection Method: Colorimetric
Research Category: Immunology
Sample Type (Specific): Supernatants, Serum
For Use With (Equipment): Microplate Reader

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.

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Manuals & protocols

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