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please find attach the filled questionnaire.
thank you very much
Asked on Apr 12 2012
Thank you very much for taking the time to complete and send the questionnaire.
The details you have kindly provided will provide us with vital information for our monitoring of product quality.
Unfortunately none of these antibodies have been tested in frozen sections, and therefore they are not guaranteed in this application, nor do we have specific protocols tips to provide for it.
However, there are some suggestions that may optimise the antibody performance, if you wish to give them a try:
-I would recommend decreasing the permeabilization conditions. Both proteins are cytoplasmic; there is no need for such an aggressive permeabilization step. I would repeat the staining skipping this step and removing Triton from the blocking and the washing buffer as well as from the antibody diluent.
-Use milder blocking conditions, using 5% of serum if possible from the same species as the secondary was raised in, to avoid cross reactivity.
-Try using more concentrated dilutions, especially for Raptor antibody (1/50 – 1/25).
If possible I would recommend trying both antibodies in paraffin embedded sections, as they are both guaranteed for it. The link to our protocols page is below, where you can find all the information regarding the different applications protocols, and very useful information for performing paraffin embedded immunostaining.
Should the suggestions not improve the results, please do not hesitate to contact me again and I’ll be more than happy to help you further.
Answered on Apr 12 2012