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.(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). 2. Wash 2-3 with distilled or deionized water. 3. Incubate sections/cell smear with Endoblocker (Not supplied) for 5-10 minutes at room temperature (RT). Wash with distilled water 3X. 4. Note: If antigen retriever (Trypsin AR-6541, Pronase AR-6542, Pepsin AR-6543, Citrate buffer AR-6544, Buffer w EDTA pH 8.5 AR-6545, Tris buffer pH 10 AR-6546) 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 (with 0.02-0.05% nonionic detergent, Triton X100, Tween 20 or NP-40) or washing buffer (Immuno Automation buffer IBSC cat # AR-6561) 3X. 6. Incubate sections/ cell smear in Protein blocking solution (Not supplied), for 5-10 minutes at RT. 7. Incubate sections/cell smear with primary antibody (NOT SUPPLIED) for 20-30 minutes at 8. Wash slide with PBS 5-7X 9. Incubate with One-Step HRP polymer (Supplied) for 20-30 minutes at RT. 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 (Not supplied). 13. Wash 5-7X with buffer. 14. Incubate with counterstain (Not supplied). 15. Wash slide with tap water distilled water. 16. Mount slide with appropriate mounting medium.

IH-8067-110 Ready to use 110 mL = 1,100 tests when 0.1 mL is applied per slide Reagents required but not supplied: Peroxidase block, protein blocking solution, Washing buffer, antigen retrievers, positive or negative control, primary antibody, primary antibody dilution buffer, chromogen, counterstain and mounting medium

Immunohistochemistry (IHC)/Immunocytochemistry (ICC) is the localization of antigens by the use of antigens in tissue sections/cells by the use of labeled antibodies as specific reagents through antigen-antibody interactions that are visualized by a marker such as fluorescent dye, enzyme, radioactive element or colloidal gold. Several IHC techniques are commonly used: labeled biotin secondary antibody streptavidin-peroxidase, HRP anti-HRP, ABC, catalyzed signal amplification, polymer system (one or two steps) and others, to detect antigens on tissue and cells. This Polymer technology shown to provide increased sensitivity and detection. IBSC One-Step goat anti-Rabbit IgG (H+L); biotin/avidin free system stains membranes, cytoplasmic and nuclear antigens. It provides the user with a rapid and easy to use IHC detection system.