One step Polymer HRP Rabbit IgG (H+L) IHC Detection Kit

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 samples two to three times with distilled or deionized water. Incubate sections or cell smears with Endoblocker for 5-10 minutes at room temperature, followed by three washes with distilled water. If antigen retrieval is required, it can be applied at this step based on 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. Wash the slides once with PBS, though this step may be omitted. Reapply the blocking solution for an additional 5-10 minutes at room temperature. Incubate sections or cell smears with the primary antibody for 20-30 minutes at room temperature; the primary antibody dilution buffer may also serve 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, ensuring that sodium azide is avoided as it destroys peroxidase 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 finally mount the slides using the appropriate mounting medium.