Pretreatment reagents are used for blocking endogenous peroxidase activity and to optimally permeabilize properly prepared samples. These reagents have been optimized to better access target RNA and reduce optimization time in multiple tissues such as formalin-fixed, paraffin-embedded (FFPE) (including archival) tissue, fresh‑frozen (FF) tissue, fixed‑frozen tissue, tissue microarray (TMA), and cell preparations.
The pretreatment reagents listed in this section are for the manual RNAscope™ and BaseScope™ Assays. Please refer to the appropriate sample preparation guides as shown in the User Manual Selection Guide available here to ensure your samples are prepared correctly for the RNAscope and BaseScope detection assay.
RNAscope 2.5 Universal Pretreatment Reagents (322380): this kit includes all the pretreatment reagents that may be used across different tissue types:
- RNAscope Target Retrieval Reagent
- RNAscope Hydrogen Peroxide Reagent
- RNAscope Protease III Reagent*
- RNAscope Protease IV Reagent*
- RNAscope Protease Plus Reagent*
The BaseScope Reagent kit is shipped with the Universal Pretreatment kit.
Select the pretreatment reagents you need based on the sample type and detection assay type:
| Tissue Type | Detection Assay Type | Reagent to use for pretreatment |
|---|---|---|
| FFPE (Formalin-Fixed, Paraffin-Embedded ) | RNAscope 2.5 HD Brown, Red, Duplex | RNAscope Hydrogen Peroxide RNAscope Target Retrieval RNAscope Protease Plus* |
| RNAscope Multiplex Fluorescent v2 | RNAscope Hydrogen Peroxide RNAscope Target Retrieval RNAscope Protease III | |
| BaseScope Red | RNAscope Hydrogen Peroxide RNAscope Target Retrieval RNAscope Protease III | |
| Fixed Frozen | RNAscope 2.5 HD Brown, Red, Duplex | RNAscope Hydrogen Peroxide RNAscope Target Retrieval RNAscope Protease Plus |
| RNAscope Fluorescent Multiplex | RNAscope Target Retrieval RNAscope Protease III | |
| RNAscope Multiplex Fluorescent v2 | RNAscope Hydrogen Peroxide RNAscope Target Retrieval RNAscope Protease III | |
| Fresh frozen | RNAscope 2.5 HD Brown, Red, Duplex | RNAscope Hydrogen Peroxide RNAscope Protease IV* |
| RNAscope Fluorescent Multiplex | RNAscope Protease IV | |
| RNAscope Multiplex Fluorescent v2 | RNAscope Hydrogen Peroxide RNAscope Protease IV | |
| BaseScope Red | RNAscope Hydrogen Peroxide RNAscope Protease IV | |
| Cultured Adherent cells | RNAscope 2.5 HD Brown, Red, Duplex | RNAscope Hydrogen Peroxide RNAscope Protease III |
| RNAscope Fluorescent Multiplex | RNAscope Protease III | |
| PBMC (Peripheral Blood Mononuclear Cells)/Non-Adherent Cells | RNAscope Fluorescent Multiplex | RNAscope Protease III |
*The protease concentration across RNAscope™ Protease Plus, Protease III and Protease IV is: Protease IV (strong conc.) > Protease III (standard)> Protease Plus (mild)
Product Ordering Information
322000 RNAscope Target Retrieval Reagents
322340 RNAscope Protease III & Protease IV Reagents
322330 RNAscope H202 & Protease Plus Reagents
322380 RNAscope 2.5 Universal Pretreatment Reagents.
