Overview

  • Product name

    Rat VEGF ELISA Kit
    See all VEGFA kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 2 pg/ml
  • Range

    0.82 pg/ml - 200 pg/ml
  • Recovery

    97 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98.26 84% - 103%
    Serum 97.43 83% - 103%
    Plasma 96.42 85% - 104%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Rat
  • Product overview

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

    This assay employs an antibody specific for rat 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-rat 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab100786 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 rat spleen and rat bone marrow showing quantity (pg) per mL of tested sample. Samples were diluted 9-27 fold.

  • VEGF measured in A10 cell culture supernatant and biological fluids showing quantity (pg) per mL of tested sample. RBL-1, PC12 and SV40LT-SIV cell culture supernatants were also assayed for rat VEGF and measured > 200 pg/mL.

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

Protocols

References

This product has been referenced in:

  • Tani N  et al. Prolactin selectively transported to cerebrospinal fluid from blood under hypoxic/ischemic conditions. PLoS One 13:e0198673 (2018). Read more (PubMed: 29949606) »
  • Nathan J  et al. Fibrocartilage Stem Cells Engraft and Self-Organize into Vascularized Bone. J Dent Res 97:329-337 (2018). Read more (PubMed: 29020504) »
See all 8 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abreviews
good ELISA kit

Abcam user community

Verified customer

Submitted Jan 24 2018

Question


First of all, I want to thank you for your immediate and explicit explanations.
I have requested the purchase date and order number of ab100786 VEGF
and the others from the university. I will email them with a better
resolution of the results to you ASAP I get access to them.
For the TNF kit ab46070, as I have understood, it seems like nothing
more can be done with the results. Please correct me if I am wrong.
For the third kit, I am so so sorry, since a little misspelling I
have done. The kit number is ab100772 IL6 rat, instead of 100722 !
However, the file you have sent to me is not what I was expecting. I
do have the brochure of the kit. My problem was something else. The
standards did not work and so a curve was not drawn. I wanted you to
email me, if possible offcourse, the optimized numbers for STANDARDs
of this kit in your lab. I mean, I am aware that each kit&apos;s standards should be optimized in case, but since my standards did not give me concentrations, although I have OD of my samples, I cannot know the concentrations of IL6 in the samples. So if possible, could you provide me the optimized numbers for OD and concentrations of
standards of this kit? (I mean the raw numbers, regardless of the fact
that they should be assessed in each case separately). So I would be
able to draw a curve by them and calculate the concentration of IL6 in
my samples upon the OD that the plate reader has provided.
Here, I would like to express my gratitudes for you co-operation.
Kind Regards,
Afshin Azimirad.

Read More
Answer

Thank you for your reply.

I look forward to receiving the resuls and order number for the ab100786 VEGF rat ELISA kit.

For the TNF kit ab46070, I think it is likely to be the sample that is the problem.However, I would be happy to investigate this further. In order to do so, I would appreciate if you are able to please complete the questionaire below and also provide the order number, lot number and a copy of the results (including the standard curve). As discussed in the previous email, it is difficult to make suggestions without haveing furhter details, and so this information will be very helpful for our investigations for you. The kit is covered by our 6 month guarantee.

With regards to ab100772IL6 Rat ELISA Kit, the standard curve using the mouse standard provided with the kit shouldbe working and we can provide our guarantee for this.I am forwarding the protocol file for this kit. I am sending this to confirm that the protocol should be undertaken exactly as described in the protocol provided with the kit,including thepreparation and detectionof the standard curve as well as the samples. Could you confirm this protocol has been followed?

https://www.abcam.com/ps/products/100/ab100772/documents/ab100772-IL6-Rat-ELISA-Kit-ab100772-protocol-plain-v2.pdf

I would appreciate if you are able to provide further information and theresults of the standard from this kit (ab100722) and I will be pleased to investigate further. In order to do so, I have attached a copy of a questionaire below. This will help you to put together the information we require quickly and easily.

Thank you for your continued time and cooperation. I look forward to hearing from you.



A. General Information
- Catalogue number:


- Lot number:

- Purchase order number/ order date or preferably the Abcam order number:

- How was the kit stored?


B. Description of the problem
- Are the signals you obtained with control or standard satisfactory? (Yes/No);


- Are the sample readings as expected or inconsistent?


- Are the blank readings as expected?

- Other (e.g. missing components)


C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.



D. Sample
- Sample Type (e.g. Cell culture supernatant, Cell lysates, Tissue lysates or Serum samples)


- Sample Species (e.g. human, mouse or rat)


- Storage (at what temperature and for how long were the samples stored?)

- Sample treatment, if any



- Sometime proteins in samples interfere with readings. Have you removed proteins?


- Sample preparation


E. Protocol used:
Did you follow the protocol exactly as given on the datasheet?


F. Optimization attempts (problem solving); please describe any deviations from the protocol.



G. Have you used the same kit successfully before?


H. Do you obtain the same results every time?


I. Additional Notes; (any additional information you would like to provide)



J. Results:

Read More

Answer

Thank you for your message. I look forward to receiving the further information regarding ab100786 VEGF Rat ELISA Kitso we can investigate this further for you.

I am sorry to hear you also have some concerns regarding two other kits. I would like to reassure you that these will also be covered by the 6 month guarantee.

ab46070 TNF-alpha rat: if the standards were ok,this indicates that the sample does not contain enough TNF alpha for detection (perhaps it has become too dilute with the washing). Do you have a good positive control sample you could try? It is possible, although notas likely, that the kit is not compatible with this sample type, I am sorry this has not been tested. For our records, I would appreciate if you are able to provide the order number and date of purchase.

ab100722 kit for IL6: I am sorry this has been tested in mouse, and not rat. I am sorry we would therefore not be able to guarantee the results from rat samples. However, the standard curve using the mouse standard provided with the kit should still be working and we can provide our guarantee for this. The protocol should be undertaken exactly as described in the protocol provided with the kit. Coud you confirm this?

https://www.abcam.com/ps/products/100/ab100722/documents/ab100722-Mouse-MCP1-ELISA-Kit-ab100722-protocol-plain-v2.pdf

I would appreciate if you are able to provide further information and the results of the standard from this kit (ab100722) and I will be pleased to investigate further. In order todo so, I have attached a copy of a questionaire below. This will help you to put together the information we require quickly andeasily.

Thank you for your time and continued cooperation. I look forward to hearing from you with the requested information.




A. General Information
- Catalogue number:

- Lot number:

- Purchase order number/ order date or preferably the Abcam order number:

- How was the kit stored?


B. Description of the problem
- Are the signals you obtained with control or standard satisfactory? (Yes/No);


- Are the sample readings as expected or inconsistent?


- Are the blank readings as expected?

- Other (e.g. missing components)


C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.



D. Sample
- Sample Type (e.g. Cell culture supernatant, Cell lysates, Tissue lysates or Serum samples)


- Sample Species (e.g. human, mouse or rat)


- Storage (at what temperature and for how long were the samples stored?)

- Sample treatment, if any



- Sometime proteins in samples interfere with readings. Have you removed proteins?


- Sample preparation


E. Protocol used:
Did you follow the protocol exactly as given on the datasheet?


F. Optimization attempts (problem solving); please describe any deviations from the protocol.



G. Have you used the same kit successfully before?


H. Do you obtain the same results every time?


I. Additional Notes; (any additional information you would like to provide)



J. Results:

Read More

Answer

Thank you for your reply and for kindly providing the information.
I am sorry to confirm that the data in the attachments you have sent are not readable. The resolution of the image is not high enough to read the numbers properly, and it is faint. I would appreciate if you are able to try sending this again.

I also look forward to hearing from you with the order number.

Regarding the reading of the plate, if there is an annomoly such as a well with high reading whena plateisread once,I would suggest it would be important to wipe the plate once more and try readingit again just in case. Was the well checked to ensure this did not contain any spilt liquid?

I hope this will be helpful and I look forward to hearing back from you with the requested information.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have some concerns regarding the results from ab100786 VEGF Rat ELISA Kit.

I would like to reassure you that thiskit is covered by our 6 month guarantee. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I would appreciate if you are able to provide the following information.

1. Please provide the order number and date of purchase.

2. I would appreciate if you are able to provide an attached copy of the data from the plate reader. This will help us to assess the problem.

3. Could you confirm how many times was the plate read? Was the bottom of the plate wiped to clean it, and then re-read? Was the well checked to ensure it did not contain any liquid?

4. Is the plate reader working well reading results from other plates?

5. How has the plate been stored?

Thank you for your time and cooperation. I look forward to receiving the requested information.

Read More

Answer

Thank you for your enquiry.

I am sorry to hear that you are experiencing difficulties with this kit. Can I confirm that you are using the protocol booklet information that comes with each new kit?

This is because there have been some concentration changes to HRP-Streptavidin Concentrate (Item G) as below:

Before 2012 Mac 15th: HRP-Streptavidin Concentrate (Item G): 8ul 6000x concentrated HRPconjugated streptavidin


After 2012 Mac 15th: HRP-Streptavidin Concentrate (Item G): 200ul 120x concentrated HRP conjugated streptavidin


If you are using a previous version of the protocol booklet, then it may be that you have been over diluting the HRPconjugated streptavidin, which is why the OD value is low. If you are using the new protocol booklet, then I will be more than happy to offer you a free of charge replacement as discussed earlier. Please let me know how you would like to proceed with this enquiry.

Read More

Question
Answer

Thank you for contacting us.

First, I want to apologize to you for the delay in gathering this information. I hope this did not cause you any inconveniences.

In regards to using rat tissues in our SCF Mouse ELISA kit, there is no cross reactivity data testing this mouse kit (ab100740) with rat samples so we do not know. However, we do know that the antibody pair is binding somewhere on the extracellular domain of the SCF protein and that mouse and rat share ˜93% homology on the extracellular domain. Therefore there is a good chance that there will be cross reactivity. Nevertheless, the only way to be certain would be to test.



As to your questions about using urine with ab100786 and ab119602.

Ab119602 will work with urine future datasheets will include this.

While ab100786 was not validated with urine samples (only serum, plasma, and cell culture supernatants) these types of kit are very accommodating to many types of samples so I see no reason why urine or any other body fluid for that matter would not be compatible with this kit.



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

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