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
Educational Resources
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