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Hope this e-mail finds you well!
Please find below a complaint from one of our customerto whom we suppliedthese Abcam antibodies.
The technical formis alsoattached in this mail.
Request you to kindly look into it & adviseus accordingly todo further needful on it.
Looking forward towards your valued reply/ comments on it.
Thankingyou in advance.
With Best Regards,
Asked on Sep 21 2012
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.
Answered on Sep 21 2012