Replaces Prod. #: ADI-950-241
The Streptavidin Blocker/Diluent can be used to prevent non-specific binding in biotin-conjugated detection systems for IHC and ISH. It can also be used as a diluent for biotinylated antibodies.
This product is intended for qualitative immunohistochemistry and in situ hybridization with formalin-fixed paraffin-embedded tissue sections and frozen sections. The blocker/diluent may also be used with blood smears, cytosmears, and cell preparations.
Product Details
| Applications: | IHC
ISH (in situ hybridization)
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| Shipping: | Blue Ice Not Frozen |
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| Long Term Storage: | +4°C |
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| Technical Info/Product Notes: | The Streptavidin Blocker/Diluent is a unique blocking solution used to prevent non-specific background staining. In addition to immunohistochemistry, the Streptavidin Blocker/Diluent can also be used for ISH.
The Streptavidin Blocker/Diluent can also be used as a diluent for SAVIEW® PLUS HRP reagent (ENZ-ACC102) and SAVIEW® PLUS AP reagent (ENZ-ACC111). It can also serve as a diluent for biotinylated antibodies. |
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| Protocol: | Recommended protocol for IHC (Immunohistochemistry)
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Arrange formalin-fixed paraffin-embedded (FFPE) tissue sections in a slide holder.
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Heat in a drying oven at 55-60°C for 20 minutes.
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Deparaffinize with Xylene (or Xylene substitute) for 10 minutes.
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Follow with 100% ethanol for 6 minutes.
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Rehydrate slides for 1 minute in each step: 90%, 70%, 50% ethanol and finally in distilled or deionized water.
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Retrieve Antigens with Antigen Retrieval Reagent (Citrate, pH 6) or Antigen Retrieval Reagent (Tris-EDTA, pH 9) for 20 minutes at 99°C.
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Wash for 5 minutes with PBST.
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Wipe excess liquid around the section on the glass slide and encircle the tissue section with a PAP pen.
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Dry slide for 10 seconds
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Incubate the tissue to peroxidase block for 5 minutes.
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Wash with PBST for 2 minutes.
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Researcher needs to optimize the concentrations and incubation times for biotinylated primary antibodies (recommended time: 20 to 30 minutes)
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Wash with PBST for 5 minutes.
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If unlabeled mouse primary antibodies are used, sections must be incubated with biotinylated anti-mouse antibody and if unlabeled rabbit primary antibodies are used, use biotinylated rabbit anti-mouse linker (diluted in blocker/diluent) for 15 minutes.
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Wash with PBST for 5 minutes.
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Incubate sections with enough Streptavidin Blocker/Diluent to cover the tissue section for 10 minutes to block non-specific binding of SAVIEW®PLUS HRP (used for detection with chromogen)
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Use SAVIEW® PLUS Detection reagents and HIGHDEF® Chromogens to visualize antigens.
Recommended Protocol for ISH (In Situ Hybridization)
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Arrange formalin-fixed paraffin-embedded (FFPE) tissue sections in a slide holder.
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Heat in a drying oven at 55-60°C for 20 minutes.
-
Deparaffinize with Xylene (or Xylene substitute) for 10 minutes.
-
Follow with 100% ethanol for 6 minutes.
-
Rehydrate slides for 1 minute in each step: 90%, 70%, 50% ethanol and finally in distilled or deionized water.
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Wipe excess liquid around the section on the glass slide and encircle the tissue section with a PAP pen.
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Dry slide for 10 seconds.
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Expose the tissue to peroxidase block for 5 minutes.
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Wash with washing buffer for 2 minutes.
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Place the slide on a 37°C warm plate.
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Add the optimal concentration of Proteinase K (recommended: 25-80μg/mL, varies with tissue type and thickness) and incubate for 10 minutes at 37°C.
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Wash with PBST for 5 minutes.
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Researcher needs to optimize the concentrations and incubation times for biotinylated RNA or DNA probes (recommended: 10 minutes of denaturation at 95°C and at least 120 minutes for hybridization, typically at 37°C for DNA probes, and 42°C for RNA probes).
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Add denatured probe to the section.
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Cover with cover slip
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Perform stringent wash after hybridization, with 40% Formamide + 6X SSPE buffer for 10 minutes.
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Wash with PBST for 5 minutes.
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Incubate sections with enough Streptavidin Blocker/Diluent to cover the tissue section for 10 minutes to block non-specific binding of SAVIEW®PLUS Detection reagents.
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Use SAVIEW® PLUS Detection reagents and HIGHDEF® Chromogens to visualize antigens.
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| Regulatory Status: | RUO - Research Use Only |
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