I recently stained with a rabbit antibody and tyramide boost, and now want to add another rabbit anitbody requiring a tyramide boost (different secondary channel). I am using a microwave treatment for the same species staining issue, do I then need to follow up with another pressure cooker antigen retrieval? Or can I go straight into the H2O2 blocking followed by serum blocking?
Hello Clara, if one of you primary antibodies needs Tyramid boost and AR and one only Tyramide boost, the first treat your sections with the antibody without AR and then in the second step the antibody with AR. If both primary ab need Tyramide boost and AR, then you need only once a time with AR. After you have finished the first sequence you can continue directly with the blocking step (H2O2 etc) for the second sequence.