i have been doing immunohistochemistry for eight months.i am using polyclonal antibody as primary,we isolated it from rabbits.at first i got nice results with non-positive tissues. the problem was false positive staining,which i reduced thru blocking with normal, goat sera,as we get DAKO secondary antibody from goat.
now for the last four months i am not even getting a faint color on strictly positive tissues.i have tried to reduce the blocking,no results, used fresh antibody ,secondary antibody incubation time also increased,also without any results.please help .
Immunohistochemistry - no positive staining
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