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Streptavidin Blocker/Diluent

Ready-to-use blocking buffer and diluent, for use with SAVIEW® PLUS detection reagents
 
ENZ-ACC109-0100 100 ml Inquire for pricing
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Replaces Prod. #: ADI-950-241

Available from stock, Ready for shipment.
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 Specification

Applications:IHC
ISH (in situ hybridization)
 
Shipping:Blue Ice Not Frozen
 
Long Term Storage:+4°C
 
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.
 
Protocol:Recommended protocol for IHC (Immunohistochemistry)
  1. Arrange formalin-fixed paraffin-embedded (FFPE) tissue sections in a slide holder.
  2. Heat in a drying oven at 55-60°C for 20 minutes.
  3. Deparaffinize with Xylene (or Xylene substitute) for 10 minutes.
  4. Follow with 100% ethanol for 6 minutes.
  5. Rehydrate slides for 1 minute in each step: 90%, 70%, 50% ethanol and finally in distilled or deionized water.
  6. Retrieve Antigens with Antigen Retrieval Reagent (Citrate, pH 6) or Antigen Retrieval Reagent (Tris-EDTA, pH 9) for 20 minutes at 99°C.
  7. Wash for 5 minutes with PBST.
  8. Wipe excess liquid around the section on the glass slide and encircle the tissue section with a PAP pen.
  9. Dry slide for 10 seconds
  10. Incubate the tissue to peroxidase block for 5 minutes.
  11. Wash with PBST for 2 minutes.
  12. Researcher needs to optimize the concentrations and incubation times for biotinylated primary antibodies (recommended time: 20 to 30 minutes)
  13. Wash with PBST for 5 minutes.
  14. 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.
  15. Wash with PBST for 5 minutes.
  16. 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)
  17. Use SAVIEW® PLUS Detection reagents and HIGHDEF® Chromogens to visualize antigens.

Recommended Protocol for ISH (In Situ Hybridization)
  1. Arrange formalin-fixed paraffin-embedded (FFPE) tissue sections in a slide holder.
  2. Heat in a drying oven at 55-60°C for 20 minutes.
  3. Deparaffinize with Xylene (or Xylene substitute) for 10 minutes.
  4. Follow with 100% ethanol for 6 minutes.
  5. Rehydrate slides for 1 minute in each step: 90%, 70%, 50% ethanol and finally in distilled or deionized water.
  6. Wipe excess liquid around the section on the glass slide and encircle the tissue section with a PAP pen.
  7. Dry slide for 10 seconds.
  8. Expose the tissue to peroxidase block for 5 minutes.
  9. Wash with washing buffer for 2 minutes.
  10. Place the slide on a 37°C warm plate.
  11. 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.
  12. Wash with PBST for 5 minutes.
  13. 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).
  14. Add denatured probe to the section.
  15. Cover with cover slip
  16. Perform stringent wash after hybridization, with 40% Formamide + 6X SSPE buffer for 10 minutes.
  17. Wash with PBST for 5 minutes.
  18. 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.
  19. Use SAVIEW® PLUS Detection reagents and HIGHDEF® Chromogens to visualize antigens.
 

Related Products

SAVIEW® PLUS HRP reagent 

High sensitivity, low background streptavidin-based nanopolymer detection reagent for use with HIGHDEF® chromogens in ISH and IHC applications
IHC, ISH (in situ hybridization) | Print as PDF
 
ENZ-ACC102-0150 150 tests Inquire for pricing
Do you need bulk/larger quantities?
 

SAVIEW® PLUS AP Reagent 

High sensitivity, low background, streptavidin-based nanopolymer detection reagent for use with HIGHDEF® chromogens in ISH and IHC applications
IHC, ISH (in situ hybridization) | Print as PDF
 
ENZ-ACC111-0150 150 tests Inquire for pricing
Do you need bulk/larger quantities?
 

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