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Here is my answer.
I am looking forward to hearing from you soon.
Asked on Nov 13 2012
Thank you for sending the details.
I am sorry without an image it is difficult to understand what is going on with staining. From description of the problem I can sense that the polyclonal antibody might be giving non-specific staining or high background - this is common with polyclonal antibodies as they need more optimization than monoclonal products. Also in double immunostaining one of the requirement is to use the individually optimized antibodies so I would strongly suggest to try these antibodies individually and observing the results. Once the satisfactory results are achieved, double immunostaining can then be performed using same conditions.
The immunohistochemistry on paraffin embedded sections is quite tricky,, in most cases antigen retrieval method needs further optimization.
It will also be good to verify the specificity of secondary antibodies to their respective primaries.
We also haven't tried these antibodies in double immunostaining so the conditions has to be determined by end user.
I hope these suggestions will be helpful. Please let me know if there is no improvements we will be happy to assist further.
Answered on Nov 13 2012