Opal Secondary Antibodies

Overview

Whether you are working on human tissue with mouse and rabbit primary antibodies or mouse tissue with rabbit primary antibodies, Opal Polymer Secondary Antibodies offer the sensitivity and specificity to make each multiplex assay a success.

Catalog

Details

1X Opal Anti-Ms + Rb HRP

1X Opal Anti-Ms + Rb HRP is a ready-to-use immunohistochemistry detection reagent specifically optimized for use with Opal and TSA^® Plus detection reagents for maximum sensitivity, reduced incubation time and low background. This product reacts specifically with primary antibodies raised in mouse and rabbit and is part of a rapid, biotin-free detection protocol. Quantitative results may be obtained with careful optimization of the primary antibody concentration. Size: 50 mL

• One solution for all mouse and rabbit primary antibodies
• Highest sensitivity 10 minute incubation
• Ready to use, dropper bottle
• Biotin-free protocol

Opal Anti-Rb HRP Kit

Opal Polymer Anti-Rabbit HRP Kit is a concentrated immunohistochemistry (IHC) detection reagent specifically optimized for use with Opal fluorescent detection reagents, providing maximum sensitivity, reduced incubation time, and low background on mouse or human FFPE tissue. This product reacts specifically with primary antibodies raised in rabbit and is part of a rapid, biotin-free detection protocol. Quantitative results may be obtained with careful optimization of the primary antibody concentration. This kit contains Opal Polymer Anti-Rabbit HRP and Opal Polymer Anti-Rabbit HRP Diluent. Recommended working dilution is 1:5.

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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