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The human VEGF ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human vascular endothelial growth factor (VEGF) with a sensitivity of 2.7 pg/mL. The assay employs a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human VEGF sandwich ELISA kit:- Colorimetric assay with a 450 nm readout; compatible with any standard plate reader- Available in different formats to meet various throughput needs, including 384-well plates for higher throughput- Broad sample compatibility – suitable for serum, plasma (heparin, citrate, EDTA), cell culture supernatants, cell culture extracts, cerebrospinal fluid, saliva, urine, and milk- Cited in over 30 publications


Images

Sandwich ELISA - Human VEGF ELISA Kit (AB222510), expandable thumbnail
  • Sandwich ELISA - Human VEGF ELISA Kit (AB222510), expandable thumbnail
  • Sandwich ELISA - Human VEGF ELISA Kit (AB222510), expandable thumbnail
  • Sandwich ELISA - Human VEGF ELISA Kit (AB222510), expandable thumbnail
  • Sandwich ELISA - Human VEGF ELISA Kit (AB222510), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Saliva, Urine, Cell culture extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

12.5 - 800 pg/mL

Assay time

1h 30m

Sensitivity

= 2.7 pg/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info
-
Notes

-

Associated Products

Select an associated product type

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Target data

Function

N-VEGFParticipates in the induction of key genes involved in the response to hypoxia and in the induction of angiogenesis such as HIF1A (PubMed:35455969). Involved in protecting cells from hypoxia-mediated cell death (By similarity).VEGFAGrowth factor active in angiogenesis, vasculogenesis and endothelial cell growth (PubMed:34530889). 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. Binds to the NRP1/neuropilin-1 receptor. Binding to NRP1 initiates a signaling pathway needed for motor neuron axon guidance and cell body migration, including for the caudal migration of facial motor neurons from rhombomere 4 to rhombomere 6 during embryonic development (By similarity). Also binds the DEAR/FBXW7-AS1 receptor (PubMed:17446437).Isoform VEGF165BBinds to the KDR receptor but does not activate downstream signaling pathways, does not activate angiogenesis and inhibits tumor growth.

Alternative names

What's included?

1 x 384 Tests
Components
10X Human VEGF Capture Antibody
1 x 600 µL
10X Human VEGF Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
384 well CaptSure™ microplates
1 x 1 Unit
50X Cell Extraction Enhancer Solution (ab193971)
11 x 1 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 50 mL
Antibody Diluent 4BI
1 x 6 mL
Human VEGF Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
2 x 50 mL
Stop Solution
2 x 12 mL
TMB Development Solution
2 x 12 mL

Recommended products

The human VEGF ELISA kit is a single-wash, 90-minute SimpleStep ELISA® assay designed for the quantification of human vascular endothelial growth factor (VEGF) with a sensitivity of 2.7 pg/mL. The assay employs a simple Mix-Wash-Read protocol with just one incubation and wash step.Easy-to-use, rapid, sensitive human VEGF sandwich ELISA kit:- Colorimetric assay with a 450 nm readout; compatible with any standard plate reader- Available in different formats to meet various throughput needs, including 384-well plates for higher throughput- Broad sample compatibility – suitable for serum, plasma (heparin, citrate, EDTA), cell culture supernatants, cell culture extracts, cerebrospinal fluid, saliva, urine, and milk- Cited in over 30 publications

Key facts

Detection method

Colorimetric

Sample types

Cerebral Spinal Fluid, Saliva, Urine, Cell culture extracts, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

12.5 - 800 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Sensitivity

= 2.7 pg/mL

Precision

Intra assay

Sample

Supernatant

n
5
mean
-
SD
-
C.V.

5.4

Inter assay

Sample

Supernatant

n
3
mean
-
SD
-
C.V.

5.5

Recovery

Sample specific recovery

Sample type

Cell culture extracts

Average %

= 89

Range

80 - 94 %

Sample type

Cell culture media

Average %

= 86

Range

81 - 89 %

Sample type

Cerebral Spinal Fluid

Average %

= 92

Range

89 - 95 %

Sample type

Citrate plasma

Average %

= 101

Range

95 - 107 %

Sample type

EDTA Plasma

Average %

= 98

Range

95 - 99 %

Sample type

Heparin Plasma

Average %

= 89

Range

84 - 92 %

Sample type

Milk

Average %

= 95

Range

91 - 98 %

Sample type

Saliva

Average %

= 109

Range

105 - 118 %

Sample type

Serum

Average %

= 109

Range

98 - 116 %

Sample type

Urine

Average %

= 95

Range

91 - 102 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Human VEGF ELISA Kit (ab222510) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of VEGF protein in edta plasma, hep plasma, saliva, serum, urine, cell culture extracts, milk, cell culture supernatant, and cit plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human VEGF with 2.7 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

INTERFERENCE

Serial dilutions of recombinant human VEGFR1, VEGFR2, and VEGFR3 were prepared starting at 4 ng/mL and tested for interference. No interference was observed.

SPECIES REACTIVITY

This kit recognizes human VEGF protein.

CALIBRATION

This immunoassay is calibrated against a highly purified human VEGF. The NIBSC/WHO unclassified purified human VEGF preparation 02/286 was evaluated in this kit.

The dose response curve of the unclassified standard 02/286 parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep human VEGF kit to approximate NIBSC 02/286 units, use the equation below.

NIBSC 02/286 approximate value (IU/mL) = 0.0011 x SimpleStep human VEGF value (pg/mL).

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.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Vascular endothelial growth factor A (VEGFA) also known as VEGF or vascular permeability factor (VPF) is a protein that plays a central role in angiogenesis as well as in the growth of blood vessels. The molecular weight of VEGFA varies depending on the isoform typically ranging from 34 to 42 kDa. VEGFA is expressed in many tissues including fibroblasts macrophages endothelial cells and platelets. Its expression levels can increase in response to stimuli like hypoxia or inflammatory cytokines. VEGFA is an important target in research for therapies related to vascular diseases and cancer.

Biological function summary

VEGFA functions by promoting the proliferation and migration of vascular endothelial cells. It operates both as a homodimer and as part of complex signaling cascades interacting with VEGF receptors on cell surfaces to trigger downstream signals for angiogenesis. This includes endothelial cell growth migration and the new blood vessel formation process. VEGFA's activity is important for physiological processes such as wound healing and embryonic development and it also contributes to pathological conditions through its unregulated expression in diseases.

Pathways

Many processes involve VEGFA including the PI3K/AKT pathway and the MAPK/ERK pathway. These pathways regulate cell survival growth and differentiation. Other proteins like PLGF (placenta growth factor) and VEGFB often accompany VEGFA in these pathways aiding in distinct but overlapping roles. These interactions are transmitted through binding to VEGF receptors such as VEGFR-1 and VEGFR-2 subsequently activating various signaling cascades necessary for cellular and tissue homeostasis.

Associated diseases and disorders

VEGFA relates to cancer and age-related macular degeneration among others. In cancer overexpression of VEGFA can lead to excessive angiogenesis supplying blood to tumors and facilitating tumor growth. In age-related macular degeneration upregulation of VEGFA contributes to the progression of the disease by promoting the formation of abnormal blood vessels under the retina. Anti-VEGFA therapies such as anti-VEGF antibodies (e.g. bevacizumab) can target these overexpressed pathways. VEGFA interacts with other proteins like HIF-1α which regulates its expression under hypoxia a common condition in tumors.

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21 product images

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in Sample Diluent NS + 1X Enhancer.

    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.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in 1X Cell Extraction Buffer PTR.

    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.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Interpolated concentrations of native VEGF in human serum and plasma samples.

    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).

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Interpolated concentrations of native VEGF in human breast milk (de-fatted), saliva, and urine samples.

    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.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Interpolated concentrations of native VEGF in cell culture supernatant samples.

    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).

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    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.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Serum from ten individual healthy human female donors was measured in duplicate.

    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.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Comparison of VEGF in PHA-M stimulated and unstimulated human PBMC cell culture supernatants stimulated for different durations.

    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).

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

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

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Other species reactivity was determined by measuring a 500 pg/mL protein load of various species recombinant VEGF proteins.

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

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

    Native VEGF was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS + 1X Enhancer.

    Recombinant human VEGF was spiked into the following biological samples and diluted in a 2-fold dilution series in Sample Diluent NS + 2X Enhancer and in Sample Diluent NS + 1X Enhancer, see Sample Preparation section for details.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Linearity of dilution.

    Native VEGF was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS + 2X Enhancer and in Sample Diluent NS + 1X Enhancer, see Sample Preparation section for details. Cell culture supernatants are represented by SN.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Linearity of dilution.

    Native VEGF was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS + 2X Enhancer and in Sample Diluent NS + 1X Enhancer, see Sample Preparation section for details. Cell culture supernatants are represented by SN.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Linearity of dilution.

    Native VEGF was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Assay sensitivity.

    The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in Sample Diluent NS + 1X Enhancer.

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

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed. 

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed. 

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Interpolated concentration of native VEGF was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 500 µg/mL A549 cell extract.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR. 

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Interpolated concentration of native VEGF was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 12.5% saliva.  The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS + 1X Enhancer.  

  • Sandwich ELISA - Human VEGF ELISA Kit (ab222510), expandable thumbnail

    Sandwich ELISA - Human VEGF ELISA Kit (ab222510)

    Example of human VEGF standard curve in 1X Cell Extraction Buffer PTR.

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

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