Anti-VEGF Receptor 2 antibody (ab39256)

According to our records, ab39256, ab63319, ab77258, ab86673, ab103674 was proving difficult to use in [APPL] and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Thank you for your email.

We will look forward to receive the images. On the optimization note we strongly recommend to include the blocking step in protocol; please try 5%BSA or 5-10% normal serum for 1-2 hours at room temperature with species same as secondary antibody. I am sure this step would improve the staining with ab103674 and ab39256.

I may be that the tissue sections you are using does not express the targets GCET1, ERG, CD14 or express at very low level so could you provide the details tissue type and species of tissue section tested. We are interested in knowing e.g. human brain tissue sections or house heart tissue sections etc.

I would suggest trying range of incubation with microwave e.g. for 5, 10, 15 and 20 minutes. This range would help to get the optimized incubation at which the antigen is maximum exposed.

I hope these suggestions would be helpful. Should you have any further question please do not hesitate to contact us.

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

- I am very surprised that 5 antibodies failed in row? Could you please specify which other antibody worked?
- What is the type and species of tissue used with each antibody? Could you supply staining images?
- Are the tissues paraffin fixed as mentioned in the details or paraformaldehyde fixed and paraffin embedded sections?
- What was the pH of Tris and Citrate buffer?
- Did tyou permeabilize the tissue sections by incubating them with Triton X-100.
- Metanol/ H2O2 blocking used to block the endogenous peroxidases, which gives false staining when HRP conjugated secondary is used. Have you done the blocking with Normal serum or BSA.
- What dilution of primaries were used in each experiment?
- What is the catalogue number of DAKO envision secondary antibody use? Did you use same kit for all the antibodies? What were the other secondary antibodies used?
- Have you used this detection system successfully before?
- Did you tired optimizing the antigen retrieval method?

Please note more information you will provide-resolution will be achieved quickly.

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

As requested, I have issued a free of charge replacement with the order number 1076421 for the VEGF antibody ab46154.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.