Overview

  • Product name
    Mouse VEGF ELISA Kit
    See all VEGFA kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 5 5.6%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 7.9%
  • Sample type
    Cell culture supernatant, Cell culture extracts, Tissue Extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.3 pg/ml
  • Range
    5.8 pg/ml - 370 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 109.6 102.9% - 117.1%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse
  • Product overview

    Mouse VEGF in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of VEGF protein in mouse cell culture supernatant, cell and tissue extract samples.


     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.


    Sensitivity:


    Samples diluted in Sample Diluent NS: 0.3 pg/mL


    Samples diluted in 1X Cell Extraction Buffer PTR: 0.7 pg/mL


     


     

  • 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 isoforms including 120, 164 and 188 amino acid long forms in mouse.  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.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse VEGF Capture Antibody 1 x 600µl
    10X Mouse 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 CPR 1 x 6ml
    Mouse VEGF Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 tests
    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
    • Vascular endothelial growth factor A
    • Vascular permeability factor
    • VEGF-A
    • Vegfa
    • VEGFA_HUMAN
    • VPF
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab209882 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
  • Standard curve comparison between mouse VEGF SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 9-fold increase in sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of VEGF were measured in duplicates, interpolated from the VEGFA standard curves and corrected for sample dilution. Undiluted samples are as follows: 5 µg/mL LPS-treated P388D1 Day 6 supernatant 7.5%, lung day 5 supernatant 15%, kidney day 5 supernatant 30%, heart day 5 supernatant 30%.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean VEGF concentration was determined to be 5,249 pg/mL in LPS P388D1 supernatant, 1,785 pg/mL in lung supernatant, 430 pg/mL in kidney supernatant and 30 pg/mL in heart supernatant samples.

  • 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 VEGFA concentration was determined to be 842 pg/mL in lung sample and 50 pg/mL in heart sample.

Protocols

References

ab209882 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Very speedy and easy to use, works for rat plasma

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Because of the limited availability of VEGF ELISAs against rat samples, we wanted to assess whether Abcam's Simplestep mouse VEGF ELISA works to measure VEGF in rat blood samples. The assay is very quick, the provided standards produced a very nice standard curve (r^2 = 0.9966), and it was able to detect VEGF in rat plasma neat and also diluted 1:5. However, serum diluted 1:5 did not correlate with neat concentrations. Therefore, we recommend using this kit for the detection of rat plasma VEGF.

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Verified customer

Submitted Jan 03 2017

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