Human VEGF ELISA Kit

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Cat: abs510008

Size: 96T

Human VEGF ELISA Kit

Testing Principle

The Human VEGF ELISA Kit employs a sandwich enzyme immunoassay technique that utilizes a specific anti-human VEGF antibody pre-coated on a highly sensitive plate with a high binding capacity. The test samples along with the standards and biotinylated detection antibodies are added to the wells of the plate that is enzyme-labeled. The VEGF present in the samples bind with the solid-phase antibodies and the detection antibodies to form immune complexes. The unbound material is then washed away, and horseradish peroxidase-labeled streptavidin (HRP) is added. Once again, the plate is washed, and a chromogenic substrate is added that generates a color in the dark. The reaction is then stopped by adding a stop solution, and the absorbance is determined at a wavelength of 450 nm with a reference correction wavelength of either 540 nm or 570 nm to yield accurate results.

Detection type: sandwich enzyme immunoassay

Form: pre-coated 96-well plate

Sample type: Cell culture supernatant, Serum, Plasma

Sample quantity: 100ul

Kit composition:

This microplate is designed for use in ELISA assays and consists of 96 wells made from high-quality polysyrene material. The wells are coated with capture antibody to capture proteins of interest in samples. The kit includes a standard and a detection antibody to allow for quantitative measurement of the protein concentration. A 10×reagent diluent is provided to dilute the reagents and samples for optimal assay performance. Additionally, color reagents A and B are included to generate a colorimetric signal upon binding of the detection antibody to the captured protein. A 25×wash buffer is provided to wash away unbound reagents and a stop solution is included to halt the color development reaction. To prevent evaporation and cross-contamination during incubation steps, ELISA plate sealers are provided. The kit comes with a specification sheet detailing the performance characteristics of the components.

Sensitivity:  7.8 pg / mL

Detection range:  31.2-2000 pg/mL

Recovery rate: 113-135%

Storage: 2-8 ° C

The standard curve:

Background: Human VEGF ELISA Kit

Vascular permeability factor or vascular endothelial growth factor (VEGF or VEGF-A) is a potent regulator of angiogenesis in both fetuses and adults. In the PDGF protein family, VEGF is grouped due to the presence of eight conserved cystine residues arranged in cystine junctions and dimerized through reverse parallel disulfide bonds. This unique structural feature contributes to its functionality and biological activity.

Human VEGF can be expressed in different subtypes through selective cleavage and variation in amino acid sequence length. These subtypes include VEGF121, VEGF145, VEGF165, VEGF183, VEGF189 and VEGF206, each with their own unique properties.
Amongst these subtypes, VEGF165 tends to be the most prevalent, followed by VEGF121 and VEGF189. However, only VEGF121 lacks the basic heparin-binding region and is able to spread freely throughout tissues.
It is worth noting that all the other subtypes, with their basic heparin-binding regions, tend to bind to tissues with high levels of heparin, such as the extracellular matrix, resulting in effective regulation of angiogenesis and vascular permeability.

VEGF165, which is highly homologous between humans and mice, is expressed in a wide range of cell types, including skeletal muscle cells, cardiomyocytes, hepatocytes, osteoblasts, neutrophils, macrophages, keratinocytes, brown fat cells, CD34+ stem cells, endothelial cells, and vascular smooth muscle cells, among others. Its expression is regulated by hypoxia and cytokines such as IL-1, IL-6, IL-8, tumor suppressor M, and tumor necrosis factor α. Furthermore, VEGF expression levels differ across various developmental stages and throughout adulthood.

The dimer of VEGF binds to two related tyrosine kinase receptors, which are VEGF R1 (also known as FLT-1) and VEGF R2 (FLK-1 /KDR).VEGF can also induce the homodimer and autophosphorylation of the latter. These receptors have seven extracellular immunoglobulin domains and one intracellular separate tyrosine receptor domain.Vascular endothelial cells and some other non-endothelial cells express VEGF receptors. Although VEGF and R1 have the highest affinity, VEGF R2 is the major factor in the regulation of VEGF angiogenesis. VEGF165 also binds to the brachioplatin (Semaporin) receptor, Neuropilin-1, thereby promoting complex formation with VEGF R2.

VEGF is known for its involvement in angiogenesis.During embryonic development, VEGF regulates endothelial cell proliferation, migration, and survival, and thus regulates vascular density and volume.But it doesn't play a role in the pattern of angiogenesis.VEGF promotes bone formation by recruiting osteoblasts and chondrocytes. It is also a monocyte chemokine. During postpartum, VEGF maintains the integrity of vascular endothelial cells and is an effective mitogen for large/small vascular endothelial cells.In adults, VEGF mainly plays a role in wound repair and female reproductive cycle. In diseased tissue, VEGF promotes vascular permeability.Therefore, VEGF is involved in tumor metastasis through extravasation and tumor angiogenesis.Various therapeutic strategies aimed at blocking VEGF activity are being used to control tumor angiogenesis induced by VEGF. Circulating VEGF levels are associated with the severity of autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus.

Human VEGF ELISA Kit is for research use only, not for use in diagnostic prodecures or in human.

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