One step Polymer HRP Mouse, Rabbit & Rat (H+L) Detection Kit

Procedure: IHC/ICC procedure for frozen, paraffin sections and cell smears. 1. Deparafinize and hydrate tissue sections through xylene or other clearing agents and graded alcohols. 2. Wash 2-3 with distilled or deionized water. 3. Incubate sections/cell smear with Endoblocker for 5-10 minutes at room temperature. Wash with distilled water 3X. 4. Note: If antigen retriever is required it can be applied at this step. Please refer to data sheet for the primary antibody. 5. Wash slide with PBS or Tris saline buffer or washing buffer 3X. 6. Incubate sections/ cell smear in Protein blocking solution, for 5-10 minutes at room temperature. 7. Incubate sections/cell smear with primary antibody for 20-30 minutes. 8. Wash slide with PBS 5-7X. 9. Incubate with One-Step HRP polymer for 20-30 minutes at room temperature. 10. Wash slide 5-7 times with buffer. Caution: Peroxidase reagents are destroyed by sodium azide and should be avoided in all buffers and regents. 11. Wash slide with deionized or distilled for 2-3X. 12. Incubate with AEC or DAB chromogen reagent. 13. Wash 5-7X with buffer. 14. Incubate with counterstain. 15. Wash slide with tap water distilled water. 16. Mount slide with appropriate mounting medium.