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Thank you for your help.
We have managed to get signal on initial trial IHC. However, only a very few cells are staining.
This may be due to limited expression in our tissue, but we wish to try to optimise to see if signal can be improved.
Your data sheet recommends proteinase K antigen retrieval - we used citrate buffer for the initial trial as per your general IHC notes on the Abcam website. Can you please provide exact recommended conditions for the proteinase K method?
Asked on Jan 08 2013
I am sorry for the delay in getting back to you in regards to the antigen retrieval protocol used with the anti-EGF antibody (ab9695).
Please find the protocol attached. Due to the type of tissue used in this testing of ab9695 (human skin tissue), the HEIR would not have been suitable as it can causes the skin to lift off of the slide. This does not mean that it will not be suitable with other tissue samples.
I hope this information has been of help. If I can be of any further assistance, please do not hesitate to let me know.
Answered on Jan 08 2013