Opal 4-Color IHC Kits

Opal™ is a practical workflow for the simultaneous detection of up to eight tissue biomarkers plus nuclear counterstain within a single image. The method is similar to standard immunohistochemistry (IHC) and is accessible to many laboratories where standard IHC is performed.

Catalog Number
Name | Link to Shop Online
Size
NEL810001KT Manual IHC Kit 50 slides
NEL820001KT Automated IHC Kit 50 slides

Product Details

The Opal 4-Color Manual IHC Kit contains:
  • 3 reactive fluorophores (Opal 520, Opal 570 and Opal 690)
  • Spectral DAPI
  • 1 X Plus Amplification Diluent
  • Blocking/Ab Diluent
  • Opal Polymer HRP Ms + Rb
  • AR6 Buffer
The Opal 4-Color Automation IHC Kit contains:
  • 3 reactive fluorophores (Opal 520, Opal 570 and Opal 690)
  • Spectral DAPI
  • 1 X Plus Automation Amplification Diluent
  • Blocking/Ab Diluent
  • Opal Polymer HRP Ms + Rb

Users should source primary antibodies and other workflow reagents separately. The Opal 4-Color Manual IHC Kit was optimized for use on Vectra® and Mantra™ systems. Kits contain enough reagents to stain 50-slides at the recommended 1:100 or 1:150 dilution ratio.

Specifications

Shipping Condition Blue Ice
Sample Human
Product Category Opal Kits
Type Opal Detection Kit
Detection Method Mantra and Vectra Quantitative Pathology Imaging Instruments

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

Documents

Phenoptics Portflolio
Brochure

Download

Opal Assay Development Guide
User Manual
Comprehensive user manual for Opal staining.

Download

Opal 4 Color Manual and Automated Multiplex IHC Assay
Data Sheets and Staining Manual

Download Staining Manual
Download Manual IHC Data Sheet
Download Automated IHC Data Sheet