Automatic & Manual Multiplexing Kits

Overview

The Tumor Infiltrating Lymphocyte 6-Plex Panel enables phenotyping of T cells, B cells and cancer cells within intact FFPE tissue sections. Kit is ideal for studying immune cell infiltration of tissues including tumors.

 

Catalog

Catalog #	Product	Size
OP7TL3001KT	Tumor Infiltrating Lymphocyte 6-Plex Panel	Up to 50 Slides

 

Details

The Tumor Infiltrating Lymphocyte 6-Plex Panel includes all critical workflow reagents to enable phenotyping of T cells, B cells and cancer cells within intact formalin-fixed, paraffin-embedded tissue sections using 6-plex fluorescent immunohistochemistry:

• Primary antibodies (anti-CD4, anti-CD8, anti-CD20, anti-CD45RO, anti-FoxP3 and anti-Pan-Cytokeratin
• 1X Antibody Diluent/Block
• 6 reactive fluorophores (Opal 520, Opal 540, Opal 570, Opal 620, Opal 650, and Opal 690)
• DMSO
• 10X Spectral DAPI
• 1X Plus Manual Amplification Diluent
• Secondary Antibody 1X Opal Anti-Ms + Rb HRP
• AR6 Buffer
• AR9 Buffer

This kit was configured and validated for multispectral imaging on the Mantra and PhenoImager HT Systems. It contains enough reagents for 6-plex immunostain of up to 50 slides.

• Optimized panel for assessment of TILs in intact FFPE tissue.
• Shortest path to results
• Part of the PhenoImager^® workflow for immunophenotyping of solid tumors

 

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

 

Related products

PhenoImager™ HT