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Lymphocyte 3-Plex Panel
formerly Opal 4 Lymphocyte Kit
The Lymphocyte 3-Plex Panel enables phenotyping of helper T cells, cytotoxic T cells and B cells within intact FFPE tissue sections.
Catalog #
Product
Size
Product Details
Specifications
Workflow
Literature
Education Resources
Product Details
The Lymphocyte 3-Plex Panel includes all critical workflow reagents to enable phenotyping of helper T cells, cytotoxic T cells and B cells within intact FFPE tissue sections with 3-plex fluorescent immunohistochemistry:
- Primary antibodies (anti-CD4, anti-CD8, anti-CD20)
- 1X Antibody Diluent/Block
- 3 reactive fluorophores (Opal 520, Opal 570, and Opal 690)
- DMSO
- 10X Spectral DAPI
- 1X Plus Manual Amplification Diluent
- Antibody 1X Opal Anti-Ms + Rb HRP
- AR6 Buffer
- AR9 Buffer
This kit was validated for multispectral imaging on the Mantra and PhenoImager HT Systems and should also provide results with fluorescence microscopes that include emission filters for DAPI, FITC, CY3 and CY5. It contains enough reagents for 3-plex immunostain of up to 50 slides.
- Optimized T and B lymphocyte panel for assessment of immune cells in intact FFPE tissue
- Shortest path to results
- Part of the PhenoImager® workflow for immunophenotyping of solid tumors
Specifications
| Shipping Condition | Blue Ice |
| Sample | Human |
| 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
Opal Storage and Stability
Storage, reconstitution and dilution of Opal dyes.
Technical Brief
Education Resources
