Here are a few controls you can use to ensure assay is working correctly:
oNegative control (primary antibody only)
§This slide receives 0.05% PBS-T in place of secondary antibody
oNegative control (secondary antibody only)
§This slide receives co-detection antibody diluent in place of diluted primary antibody
§If performing staining across multiple batches, include serial sections of one tissue in each batch
Parameters for some steps (e.g. antibody incubations, antigen retrieval, chromogen incubations, counterstaining) may need to be further optimized for different tissues or targets.