Human VEGF ELISA Kit (Colorimetric)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-91272

Novus Biologicals, part of Bio-Techne
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NBP1-91272
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Key Product Details

Sample Type & Volume Required Per Well

Cell culture supernatant, serum, plasma (EDTA, citrate, heparin), buffered solution (50 uL)

Sensitivity

7.9 pg/mL

Assay Range

15.6 - 1,000 pg/mL

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Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Reactivity

Human

Specificity

The assay detects both natural and recombinant human VEGF-A. Cross reactivity and interference of circulating factors of the immune system were evaluated by spiking these proteins at physiologically relevant concentrations into a human VEGF-A positive serum. There was no crossreactivity detected, notably not with human VEGF-B, VEGF-C, VEGF-D and PfGF. Interference was detected for VEGF-R1 at concentrations > 200 pg/ml, and not for VEGF-R2.

Description

The human VEGF-A ELISA is an enzyme-linked immunosorbent assay for the quantitative detection of human VEGF-A

Precision

Intra-Assay Precision (Precision within an assay) CV% < 4.30%

Inter-Assay Precision (Precision between assays) CV% < 6.20%

Recovery for Human VEGF ELISA Kit (Colorimetric)

Recovery

Please see protocol for more specific Spike Recovery details.

Scientific Data Images for Human VEGF ELISA Kit (Colorimetric)

ELISA: Human VEGF ELISA Kit (Colorimetric) [NBP1-91272]

ELISA: Human VEGF ELISA Kit (Colorimetric) [NBP1-91272]

ELISA: VEGF ELISA Kit [NBP1-91272] - Standard Curve

Kit Contents for Human VEGF ELISA Kit (Colorimetric)

  • Adhesive Films
  • Aluminium pouch(es) with a Microwell Plate coated with polyclonal antibody to human VEGF-A
  • Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
  • Biotin-Conjugate anti-human VEGF-A polyclonal antibody
  • Human VEGF-A Standard lyophilized, 2 ng/mL upon reconstitution
  • Sample Diluent
  • Stop Solution (1M Phosphoric acid)
  • Streptavidin-HRP
  • Substrate Solution (tetramethyl-benzidine)
  • Wash Buffer Concentrate 20x (PBS with 1% Tween 20)

Preparation and Storage

Formulation

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage of components varies. See protocol for specific instructions.

Background: VEGF

Vascular endothelial growth factor (VEGF), also called VEGF-A and vascular permeability factor (VPF), is a secreted homodimeric glycoprotein belonging to the VEGF family with a role in stimulating angiogenesis and vasculogenesis (1,2). More specifically, VEGF-A secretion from most cell types contributes to promoting endothelial cell proliferation and migration, inhibiting apoptosis, increasing vascular permeability, and wound healing (1). The VEGF family consists of several members including VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF-F, and placenta growth factor (PLGF) (1-4). As a result of alternative splicing of the eight exon VEGFA gene, there are several VEGF-A protein isoforms of 121, 145, 165, 183, 189, and 206 amino acids (aa) in length, with VEGF121 and VEGF165 being the two most expressed isoforms (1,5). Full length VEGF-A monomer has a 26 aa signal sequence plus a 206 aa (VEGF206) sequence, with a theoretic molecular weight (MW) of 27 kDa, containing VEGF receptor 1 (VEGFR1) and VEGR2 binding sites and heparin-binding domains (1-3,5,6). VEGF121 lacks heparin affinity and binds the receptor tyrosine kinases (RTKs) VEGFR1 and VEGFR2, whereas VEGF165 has moderate affinity for heparin and, in addition to being a ligand for VEGFR1 and VEGFR2, can also bind the co-receptors neuropilin 1 (NRP1) and NRP2 (1,5). Hypoxia and hypoxia-related genes such as HIF-1, EGF, and PDGF are major regulators angiogenesis and VEGF expression (1,3). VEGF signaling initiated by ligand binding to its receptors results in activation of different pathways including PI3K and MAPK and ultimately guides endothelial cell proliferation, migration, and survival (1,3). While VEGF plays an important role in promoting normal angiogenesis and blood vessel formation, its expression is often upregulated in tumors and other angiogenesis-related pathologies like osteroarthritis (OA) (1-5,7). Given its function, VEGF and its receptors have become a therapeutic target for treating cancer and blocking angiogenesis (4,5,7). A recombinant humanized monoclonal anti-VEGFA antibody called bevacizumab (Avastin) was first approved by the FDA in 2004 for the treatment of a number of cancers (1-3,5). Cancer patients may experience resistance to anti-VEGF antibodies and, as such, clinical studies are exploring combination treatment options with chemotherapies and immune-checkpoint inhibitors (3,5).

References

1. Melincovici CS, Bosca AB, susman S, et al. Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis. Rom J Morphol Embryol. 2018;59(2):455-467.

2. Shaik F, Cuthbert GA, Homer-Vanniasinkam S, Muench SP, Ponnambalam S, Harrison MA. Structural Basis for Vascular Endothelial Growth Factor Receptor Activation and Implications for Disease Therapy. Biomolecules. 2020;10(12):1673. https://doi.org/10.3390/biom10121673

3. Apte RS, Chen DS, Ferrara N. VEGF in Signaling and Disease: Beyond Discovery and Development. Cell. 2019;176(6):1248-1264. https://doi.org/10.1016/j.cell.2019.01.021

4. Matsumoto K, Ema M. Roles of VEGF-A signalling in development, regeneration, and tumours. J Biochem. 2014;156(1):1-10. https://doi.org/10.1093/jb/mvu031

5. Itatani Y, Kawada K, Yamamoto T, Sakai Y. Resistance to Anti-Angiogenic Therapy in Cancer-Alterations to Anti-VEGF Pathway. Int J Mol Sci. 2018;19(4):1232. Published 2018 Apr 18. doi:10.3390/ijms19041232

6. Uniprot (P15692)

7. Hamilton JL, Nagao M, Levine BR, Chen D, Olsen BR, Im HJ. Targeting VEGF and Its Receptors for the Treatment of Osteoarthritis and Associated Pain. J Bone Miner Res. 2016;31(5):911-924. https://doi.org/10.1002/jbmr.2828

Long Name

Vascular Endothelial Growth Factor

Alternate Names

MVCD1, VAS, Vasculotropin, VEGF-A, VEGFA, VPF

Gene Symbol

VEGFA

Additional VEGF Products

Product Documents for Human VEGF ELISA Kit (Colorimetric)

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Certificate of Analysis

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Product Specific Notices for Human VEGF ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

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