Overview

  • Product name
    Human VEGF ELISA Kit
    See all VEGFA kits
  • Detection method
    Colorimetric
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    < 10 pg/ml
  • Range
    8.23 pg/ml - 6000 pg/ml
  • Recovery

    > 100 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 103.5 92% - 113%
    Serum 104.4 92% - 115%
    Plasma 105.7 93% - 114%

  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s VEGF Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human VEGF in serum, plasma and cell culture supernatants.

    This assay employs an antibody specific for Human VEGF coated on a 96-well plate. Standards and samples are pipetted into the wells and VEGF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human VEGF antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of VEGF bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimization may be required with urine samples.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab100662 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

  • VEGF measured in biological fluids showing quantity (pg) per milliliter of sample tested

  • Curves showing concentrations huVEGF165 in NIBSC 02/286 using ab100662 VEGF human

     

  • Representative standard curve using ab100662

  • Representative standard curve using ab100662

Protocols

References

This product has been referenced in:
  • Tong M  et al. Micro- and Nano-vesicles from First Trimester Human Placentae Carry Flt-1 and Levels Are Increased in Severe Preeclampsia. Front Endocrinol (Lausanne) 8:174 (2017). Read more (PubMed: 28790977) »
  • Steinecker-Frohnwieser B  et al. Pharmacological treatment with diacerein combined with mechanical stimulation affects the expression of growth factors in human chondrocytes. Biochem Biophys Rep 11:154-160 (2017). Sandwich ELISA ; Human . Read more (PubMed: 28955780) »
See all 6 Publications for this product

Customer reviews and Q&As

1-10 of 14 Abreviews or Q&A

Answer



Please find below the information regarding the source/host of our capture and detection antibodies used in our VEGF Human ELISA Kit (ab100662):

Capture Antibody: Monoclonal Mouse IgG2B

Detection Antibody: Polyclonal Goat IgG

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Answer


Please note that ab100662 and ab100663 are similar kits that both measure levels of VEGF.

However ab100662 is best used for detecting VEGF in biological fluids such as cell culture supernatant, serum and plasma while ab100663 is best suited for use with cell and tissue extracts.

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Question
Answer

Unfortunately we no longer test urine as part of the validation. However the kit is very likely to work for urine (based on our past experience with this sample type).

Here are so some general guidelines on preparation of urine for ELISA testing:
- Collect urine sample without adding stabilizers.
- Centrifuge at 10,000xg for 2 min at 4°C.
- Aliquot supernatant quickly in dry ice/methanol bath and store samples at -80°C. Minimize freeze/thaw cycles.

We would also recommend starting with a minimal dilution (such as 2-fold), since in our experience, cytokine levels are often very low in urine.

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Answer



Capture Antibody: Monoclonal, mouse. The immunogen sequence is Ala27­Arg191 (Accession # NP_001165097.1) Detects human VEGF165 and human VEGF121. In ELISAs, this antibody has ˜10% cross­reactivity with recombinant mouse VEGF and rat VEGF and no cross­reactivity with recombinant human VEGF­D.



Detection Antibody: Polyclonal, goat. The immunogen sequence is Ala27­Arg191 (Accession # AAV38412). Detects human VEGF165. Showed less than 15% cross­reactivity with recombinant human (rh) VEGF/PlGF is observed and less than 0.1% cross­reactivity with recombinant mouse PlGF­2, VEGF, recombinant human Flt­1, VEGF R2, PlGF, VEGF­C, and VEGF­D.

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Question
Answer



The VEGFA isoform that ab100662 detects is 165 and 121.

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Answer



Answer 1:

Please use 1X Assay Diluent B to dilute the detection antibody and the HRP-Streptavidin. If testing serum/plasma, use Assay Diluent A to prepare/dilute the standards and samples. Where it says use the “appropriate assay diluent,” use Assay Diluent A, but when it specifically calls for the use of diluent B, use diluent B.




Answer 2:

Dilute the detection antibody concentrate, prepared in step 6 of the reagent preparation section, 100-fold with 1X Assay Diluent B. This means after adding the 100 ul of 1X Assay Diluent B to the vial, it will need to be diluted 100-fold with the same diluent (B).



I hope these answers are useful. Please let us know if you have further questions.

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Answer

Thank you for contacting us.

I can confirm that Heparin should not be interfering with the assay using these kits.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Question
Answer

Te pido disculpas por la espera.

He recibido una contestación de mis compañeros, que me comentan que la dilución recomendada para suero y plasma debe hacerse entre 2 y 5 veces.

En cualquier caso, los niveles de proteína varían entre distintas muestras y como indica el protocolo deben calcularse las diluciones óptimas para cada una.

Siento que esta información no figurara desde el principio en el protocolo, que será inmediatamente actualizado.

Te recuerdo que todos nuestros productos tienen una garantía (Abpromise) de 6 meses, y en caso de no funcionar como se especifica en su datasheet, se os reemplazará el producto de forma gratuita o se reembolsará el importe del mismo.

Para más información, no dudéis en consultarnos.

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Answer


Please note this product is unpublished so when you decide to order you need to contact our customer service team for them to put this item in your order.

I hope this info will help. Please let me know if you experience any difficulties.

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Answer

This product is available but not published on our website. You cannot purchase this using our normal ordering system however I can add this product to your next shipment. Could you confirm how many vials customer would need..6?.
In my last I made an error in calculating the price; the price would be $720 for 6 vials.

The concentration of HRP-streptavidin is not important. The 200ul solution could be used as recommended in protocol. Please note customer have to use new protocol with these new vials, which is available on our website. Each kit protocol have the information on how to use the 200ul vial of HRP-Strep conjugate.

I hope this information will be helpful.

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1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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