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Protocol for Immunohistochemistry (IHC) and Immunocytochemistry (ICC)
- If tissues are used, deparafinize and hydrate tissue sections through xylene or other clearing agents and graded alcohols. (For frozen sections or cell smears; use unfixed, acetone fixed or appropriate fixative for the antigen in question; for cell smears it may be necessary to permealize the cell by detergent, please refer to antibody protocol).
- Wash 2-3 with distilled or deionized water.
- Incubate sections/cell smear in Endoblocker (#1) for 5-10 minutes at room temperature or 37°C. Note: If antigen retriever is required it can be applied after this stage.
- Wash slide with PBS or Tris saline (with 0.02-0.05% nonionic detergent, Triton X100, Tween 20 or NP-40) or washing buffer (Immuno Automation Buffer IBSC cat # AR-6561) 3-5X.
- Incubate sections/ cell smear in Immuno Protein blocking solution (diluted animal serum), for 10 minutes at RT or 37ºC or as suggested in your protocol.
- Wash slide with PBS 1X.
- Follow instructions for IHC.