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

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 kit employs Polymer technology shown to provide increased sensitivity and detection. IBSC One-Step 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.

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-8067-15 Ready to use 15 mL = 150 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 kit employs Polymer technology shown to provide increased sensitivity and detection. IBSC One-Step 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.