1X Antibody Diluent / Block

Overview

This ready-to-use antibody blocking and diluent buffer formulation has been optimized specifically for Opal™ and TSA® Plus immunohistochemistry protocols.

Catalog

Details

This 2-in-1 antibody diluent for IHC and blocking buffer acts to minimize non-specific interaction of antibodies that can be a source of background
This formulation does not contain azide and should be compatible with all peroxidase based detection methods. Size: 100 mL.

• Ready to use formulation
• Convenient dropper bottle
• Azide-free, does not affect HRP

Workflow

Bake and Deparaffinize Slides

• Bake at 60 C for one hour
• Xylene wash (3x ten minutes)
• Rehydrate with EtOH gradient into di H2O

Slide Fixation

• 10% NBF for 20 minutes
• Di H2O wash

Antigen Retrieval

• AR6 or AR9 MWT
• Cool to RT for at least 15 minutes

Blocking

• Di H2O rinse, TBST rinse
• PAP pen barrier
• Incubate tissue for 10 minutes in blocking solution at RT

Primary Antibody

• Remove blocker and add primary antibody
• Rinse with TBST, then wash 3 times for two minutes each in TBST

Secondary Antibody

• Incubation 10 minutes at RT
• Rinse with TBST and wash 3 times for 2 minutes each

Opal Reagent

• Opal fluorophore incubation for ten minutes at RT
• Rinse with TBST and wash 3 times for 2 minutes each

MWT

• Rinse slides with AR6 or AR9
• Place slides in microwave-safe jar, perform MWT
• Cool to RT for at least 15 minutes

Spectral DAPI

• Use AR6 for MWT prior to DAPI for removal of any unbound fluor
• Rinse slides in di H2O and then TBST
• Incubate DAPI for 5 minutes at RT
• Wash with di H2O and TBST two minutes each

Mount

• Apply monunting medium for fluorescene microscopy for coverslip

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