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blocking HRP linking and HRP labeling reagents from the ultra stripped avidin HRP detection kit are used in the following sections tissue permeabilization is recommended for intracellular targets after processing and preparation incubate slides with permeabilization buffer containing 0.25 percent triton x-100 in PBS for 30 minutes at room temperature then wash in PBS three times five minutes each dry the slides carefully without disturbing the tissue apply serum blocking reagent number 2 included in the HRP detection kit two slides and incubate for a minimum of five minutes at room temperature prepare primary antibody solution in blocking buffer or antibody diluent buffer catalog number nine two six zero zero one for purified antibodies at the recommended concentration the proper diluent to use for the antibody depends on the antibodies format remove blocking buffer do not rinse apply antibody solution and incubate for an hour at room temperature incubation may vary depending o
