Help & FAQs: Staining Kits and Reagents

• My tissue stained with the VisUCyte™ HRP polymer seems to be overstained, what could I improve to avoid this?
• We would recommend lowering the concentration of the primary antibody used. Due the high sensitivity of VisUCyte™ HRP Polymer, the working concentration of primary antibodies may be 23 times lower compared to the conventional avidin-biotin-HRP protocol.
• What is the recommended procedure for reducing background on tissue sample which appears overstained after using R&D Systems VisUCyte™ HRP polymer?  
• R&D Systems recommend reducing the concentration of the primary antibody, and/or shortening the amount of time the tissue is incubated with our VisUCyte™ HRP polymer reagent.

Cell and Tissue Staining Kits

• Are deparaffinization and antigen retrieval performed prior to starting IHC detection using Cell and Tissue Staining Kits (for example, Catalog # CTS019)?
• Yes, perform deparaffinization and, if necesary, antigen retrieval prior to treating samples with primary antibody, and then use the Cell and Tissue Staining Kit for detection.

Immunohistochemistry Reagents

• Is antigen retreival required for IHC?
• Antigen retrieval is something to consider if initial IHC staining is negative.  Somtimes fixation and processing can block epitopes, and antigen retrieval can reveal these eptiopes.  For a review of antigen retreival considerations, please consult the "Resources" tab on R&D Systems' website.  Antigen retrieval information is listed under the "Protocols-Troubleshooting-Guides" link.

Staining Reagents

• My tissue stained with the VisUCyte™ HRP polymer seems to be overstained, what could I improve to avoid this?
• We would recommend lowering the concentration of the primary antibody used. Due the high sensitivity of VisUCyte™ HRP Polymer, the working concentration of primary antibodies may be 23 times lower compared to the conventional avidin-biotin-HRP protocol.
• What is the recommended procedure for reducing background on tissue sample which appears overstained after using R&D Systems VisUCyte™ HRP polymer?  
• R&D Systems recommend reducing the concentration of the primary antibody, and/or shortening the amount of time the tissue is incubated with our VisUCyte™ HRP polymer reagent.