Human VEGF ELISA Kit (ab222510)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.7 pg/ml
- Range: 12.5 pg/ml - 800 pg/ml
- Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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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
Cell culture supernatant, Saliva, Milk, Urine, Serum, Cell culture extracts, Hep Plasma, EDTA Plasma, Cit 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% Hep Plasma 89 84% - 92% EDTA Plasma 98 95% - 99% Cit plasma 101 95% - 107% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
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 stripsA 384-well SimpleStep ELISA® microplate (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).
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Notes
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. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 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
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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
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Alternative names
- Folliculostellate cell-derived growth factor
- Glioma-derived endothelial cell mitogen
- MGC70609
see all -
Database links
- Entrez Gene: 7422 Human
- Omim: 192240 Human
- SwissProt: P15692 Human
- Unigene: 73793 Human
Images
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Other - Human VEGF ELISA Kit (ab222510)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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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.
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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.
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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.
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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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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).
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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.
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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).
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Interpolated concentrations of native VEGF.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.
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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.
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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).
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Serial dilutions of recombinant human VEGF189 and VEGF121.Serial dilutions of recombinant human VEGF189 and VEGF121 were prepared and assayed in parallel with recombinant VEGF165.
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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).
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Linearity of dilution.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.
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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.
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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.
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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.
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Assay sensitivity.The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.
