Opal anti-rabbit HRP staining

AR9 & AR6 Buffer

These buffers are optimized for pH6 or pH9 antigen retrieval in manual Multiplex IHC staining.

Catalog Number
Name | Link to Shop Online
Size
AR900250ML AR9 Buffer, 10X 250 mL 250 mL
AR9001KT AR9 Buffer, 10X 4x 250 mL 4x 250 mL
AR600250ML AR6 Buffer, 10X 250 mL 250 mL
AR6001KT AR6 Buffer, 10X 4x 250 mL 4x 250 mL
AR600125ML AR6 Buffer, 10X 125 mL 125 mL

Product Details

10X buffer for use in Opal™ immunohistochemistry multiplexing protocols. This buffer was developed for use in Opal multicolor immunohistochemistry protocol in microwave treatment (MWT) steps. MWT provides antigen retrieval for formalin-fixed, paraffin embedded (FFPE) tissue, quenches endogenous peroxidase, and removes antibodies after epitopes have been labeled with Opal dyes.

Specifications

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

Workflows

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

Material Safety Data Sheets

Name
MSDS
AR6 Buffer Download
AR9 Buffer Download