Overview

  • Product name
    Human VEGF ELISA Kit
    See all VEGFA kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Supernatant 5 5.4%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 3 5.5%
  • Sample type
    Saliva, Milk, Urine, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    2.7 pg/ml
  • Range
    12.5 pg/ml - 800 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 109 105% - 118%
    Milk 95 91% - 98%
    Urine 95 91% - 102%
    Serum 109 98% - 116%
    Cell culture extracts 89 80% - 94%
    Cell culture media 86 81% - 89%
    Heparin Plasma 89 84% - 92%
    EDTA Plasma 98 95% - 99%
    Citrate Plasma 101 95% - 107%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Does not react with: Cow
  • Product overview

    VEGF in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of VEGF protein in human serum, plasma, milk, saliva, urine, cell culture supernatants, cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    VEGF is a secreted growth factor of PDGF family active in angiogenesis, vasculogenesis and endothelial cell growth both in fetus and adult. Alternative splicing produces many isoforms including major isoforms VEGF121, VEGF165 and VEGF189 in human. VEGF expression is induced by hypoxia. It is regulated by growth factors, cytokines, gonadotropins, nitric oxide, hypoglycemia and oncogenic mutations. VEGF induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. VEGF dimers bind to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, induce their homodimerization and autophosphorylation. VEGF165 and VEGF145 interact with NRP1/Neuropilin.


    Sensitivity:


    Sample diluted in Cell Extraction Buffer PTR - 2.9 pg/mL


    Sample diluted in Sample Diluent NS + 1X Enhancer - 2.7 pg/mL

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human VEGF Capture Antibody 1 x 600µl
    10X Human VEGF Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Human VEGF Lyophilized Recombinant Protein 1 x 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas
  • Function
    Growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Induces endothelial cell proliferation, promotes cell migration, inhibits apoptosis and induces permeabilization of blood vessels. Binds to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparan sulfate and heparin. NRP1/Neuropilin-1 binds isoforms VEGF-165 and VEGF-145. Isoform VEGF165B binds to KDR but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth.
  • Tissue specificity
    Isoform VEGF189, isoform VEGF165 and isoform VEGF121 are widely expressed. Isoform VEGF206 and isoform VEGF145 are not widely expressed.
  • Involvement in disease
    Defects in VEGFA are a cause of susceptibility to microvascular complications of diabetes type 1 (MVCD1) [MIM:603933]. These are pathological conditions that develop in numerous tissues and organs as a consequence of diabetes mellitus. They include diabetic retinopathy, diabetic nephropathy leading to end-stage renal disease, and diabetic neuropathy. Diabetic retinopathy remains the major cause of new-onset blindness among diabetic adults. It is characterized by vascular permeability and increased tissue ischemia and angiogenesis.
  • Sequence similarities
    Belongs to the PDGF/VEGF growth factor family.
  • Cellular localization
    Secreted. VEGF121 is acidic and freely secreted. VEGF165 is more basic, has heparin-binding properties and, although a signicant proportion remains cell-associated, most is freely secreted. VEGF189 is very basic, it is cell-associated after secretion and is bound avidly by heparin and the extracellular matrix, although it may be released as a soluble form by heparin, heparinase or plasmin.
  • Information by UniProt
  • Alternative names
    • Folliculostellate cell-derived growth factor
    • Glioma-derived endothelial cell mitogen
    • MGC70609
    • MVCD1
    • vascular endothelial growth factor
    • Vascular endothelial growth factor A
    • vascular endothelial growth factor A121
    • vascular endothelial growth factor A165
    • Vascular permeability factor
    • Vegf
    • VEGF A
    • VEGF-A
    • VEGF120
    • Vegfa
    • VEGFA_HUMAN
    • VPF
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab222510 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • The VEGF standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The VEGF standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (heparin) 50% and plasma (EDTA) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 268.9 pg/mL in neat serum, 123.9 pg/mL in neat plasma (citrate), 133.8 pg/mL in neat plasma (heparin), and 95.76 pg/mL in neat plasma (EDTA).

  • The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows: breast milk 2%, saliva 12.5%, and urine 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 21487 pg/mL in neat human breast milk (de-fatted), 3712 pg/mL in neat human saliva, and 262.0 pg/mL in neat human urine.

  • The concentrations of VEGF were measured in duplicates, interpolated from the VEGF standard curves and corrected for sample dilution. Undiluted samples are as follows: HepG2 5%, A431 5%, MDA-MB-435S 12.5%, PC-3 20%, A549 50%, and PBMC 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 5027 pg/mL in neat HepG2 supernatant, 7356 pg/mL in neat A431 supernatant (4 Day), 5918 pg/mL in neat MDA-MB-435S supernatant, 1495 pg/mL in neat PC-3 (2 Day), 603.4 pg/mL in neat A549 supernatant, and 405.3 pg/mL in neat PBMC supernatant (PHA-M, 5 Day).

  • Interpolated concentrations of native VEGF in PC-3 cell extract (1 Day), PC-3 cell extract (2 Day), HepG2 cell extract, A549 cell extract, and MDA-MB-435S cell extract samples based on a 300 μg/mL extract load. The concentrations of VEGF were measured in duplicate and interpolated from the VEGF standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 86.65 pg/mL in PC-3 cell extract (1 Day), 149.6 pg/mL in PC-3 cell extract (2 Day), 127.7 pg/mL in HepG2 cell extract, 241.7 pg/mL in A549 cell extract, and 454.5 pg/mL in MDA-MB-435S cell extract.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGF concentration was determined to be 235.0 pg/mL with a range of 81.10 – 993.3 pg/mL.

  • Human PBMC (seeded at 10x106/mL) were cultured for 2 or 5 days in the presence or absence of 1.5% PHA-M. The concentrations of VEGF were measured in three different dilutions of the supernatant samples in duplicates and interpolated from the VEGF standard curve. The interpolated values are plotted (mean +/- SD, n=3). The mean VEGF concentration was determined to be 34.76 pg/mL in 2 Day PHA-M stimulated PBMC cell culture supernatant, 393.7 pg/mL in 5 Day PHA-M stimulated PBMC cell culture supernatant, and undetectable in unstimulated PBMC cell culture supernatant at both 2 and 5 days (not shown).

  • Serial dilutions of recombinant human VEGF189 and VEGF121 were prepared and assayed in parallel with recombinant VEGF165.

  • Protein concentrations were interpolated from the human standard curve, and graphing the interpolated concentrations (mean +/- SD, n=2).

Protocols

References

This product has been referenced in:
See 1 Publication for this product

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