One Step Polymer HRP Mouse and Rat IgG (H+L)

IHC/ICC procedure for frozen, paraffin sections and cell smears begins by deparaffinizing and hydrating tissue sections through xylene or other clearing agents and graded alcohols. Wash the specimens two to three times with distilled or deionized water. Incubate sections or cell smears with Endoblocker for 5-10 minutes at room temperature, then wash three times with distilled water. If an antigen retriever is required, it can be applied at this step following the primary antibody data sheet. Wash the slides three times with PBS, Tris saline buffer, or washing buffer. Incubate sections or cell smears in Protein blocking solution for 5-10 minutes at room temperature. Incubate sections or cell smears with primary antibody for 20-30 minutes at room temperature; the primary antibody dilution buffer can also be used as a negative control. Wash the slides five to seven times with PBS. Incubate with One-Step HRP polymer for 20-30 minutes at room temperature. Wash the slides five to seven times with buffer, noting that peroxidase reagents are destroyed by sodium azide and should be avoided in all buffers and reagents. Wash the slides two to three times with deionized or distilled water. Incubate with AEC or DAB chromogen reagent, then wash five to seven times with buffer. Incubate with counterstain, wash with tap water or distilled water, and mount the slide with the appropriate mounting medium.