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1x Plus Amplification Diluents
These ready-to-use diluents have been optimized specifically for Opal™ and TSA® Plus immunohistochemistry protocols.
Product Details
Specifications
Workflow
Literature
Data Sheets
Education Resources
Product Details
Utilize these diluents to bring your Opal fluorophores to working concentration.
1X Plus Manual Amplification Diluents are for use with Opal fluorophores when staining manually, typically at 1:100 dilution. The 1X Plus Automation Amplification Diluent is formulated specifically for Opal multiplex IHC fluorophores used on the Leica BOND RX, for use at 1:150.
Specifications
Shipping Condition | Blue Ice |
Sample | Human or Mouse |
Product Category | Opal Kits |
Type | Opal Detection Kit |
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 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
Education Resources