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AR6 & AR9 Buffers
These buffers are optimized for pH6 or pH9 antigen retrieval in manual Multiplex IHC staining.
Product Details
Specifications
Workflow
Literature
Data Sheets
Education Resources
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 2, Vectra 3, PhenoImager Fusion, and PhenoImager HT |
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 microwave treatment (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
Literature
Data Sheets
Name
MSDS
AR6 Buffer (125ml)
AR6 Buffer (125ml)
AR6 Buffer (250ml)
AR6 Buffer (250ml)
AR9 Buffer (250ml)
AR9 Buffer (250ml)
Education Resources