Dear expertise,
i stained frozen section lung tissue sample against virus protein, which should be detect virus infection. i trying to reduce non-specific interaction using serum (5% BSA in PBS) blacking steps to prevent non-specific interaction. its not sufficient in order to control non-specific binding.
i need your help/suggestion to control non-specific interaction in IHC.
Thank you very much for your valuable help/reading.
-N. Stalin
I agree with Sebastian so far. I recommand to run a negativ control. That means take one of your lung sections and add instaed of the primary antobody only the delution medium. After zhe incubation continue with your protocol. If your system (secondary antibody and or StreptAvidin or other detection systems) works well the negativ control should be negativ and possible unspecific background staining comes from the primary ab. Try different dilutions to optimize the working dilution of your primary ab.
We usually use ~ 5% serum deriving from the same species as the secondary antibody (eg. normal goat serum) and BSA for blocking. You can also add gelatine (we use 0.2%) if this isn't sufficient.
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