1x Plus Amplification Diluents

These ready-to-use diluents have been optimized specifically for Opal™ and TSA® Plus immunohistochemistry protocols.

Catalog Number
Name | Link to Shop Online
size
FP1498 1X Plus Amplification Diluent 50 mL
FP1609 1X Plus Automation Amplification Diluent 50 mL

Product Details

Utilize these diluents to bring your Opal fluorophores to working concentration.

1X Plus Amplification Diluent is 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 and Vectra Quantitative Pathology Imaging Instruments

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 ten 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 for ten minutes at RT
  • Rinse with TBST and wash 3 times for two minutes each
Opal Reagent
  • Opal fluorophore incubation for ten minutes at RT
  • Rinse with TBST and wash 3 times for two 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 mounting medium for fluorescence microscopy and coverslip

Documents

Phenoptics Portfolio
Brochure

Download

Opal Assay Development Guide
User Manual
Comprehensive user manual for Opal staining.

Download

Material Safety Data Sheets

Catalog Number
Name
MSDS
FP1498 1X Plus Amplification Diluent Download
FP1609 1X Plus Automation Amplification Diluent Downlaod

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