The RNAscope 2.5 HD Duplex Assay is based on ACD’s patented signal amplification and background suppression technology. The assay uses a novel and proprietary method of in situ hybridization (ISH) to visualize single RNA molecules per cell in FFPE samples mounted on slides. RNAscope Assays do not require the RNA-free environment used for traditional ISH. RNAscope probes in Channel 1 and Channel 2 are compatible with this assay which utilizes HRP-based Green and AP-based Fast Red chromogens to result in a detectable green and red signal respectively. The RNAscope 2.5 HD Duplex Assay can be used for simultaneous visualization of two RNA targets while maintaining single cell resolution. The applications of this assay include study of gene co-regulation, profile gene expression in a specific cell type (identified using a cell specific marker) and map coexpression of two targets within the same cellular context based on reliable chromogenic detection of the two transcripts.
RNAscope™ 2.5 HD Duplex Assay
RNAscope 2.5 HD Duplex Reagent Kit gives reliable results which are ideal for archiving due to the permanent staining visible under any standard bright field microscope. The RNAscope 2.5 HD Duplex Reagent Kits are designed to provide more intense staining when low copy target gene expression is anticipated (1–20 copies per cell). The RNAscope 2.5 HD Duplex reagent kits are compatible with the same RNAscope probes used previously with RNAscope 2.0 2-plex assays.
For customers using RNAscope 2.5 HD Duplex Manual Assay for the first time, ACD's Intro Packs include species-specific control slides, control probes, the Reagent Kit, the ImmEdge hydrophobic barrier pen and the must-have accessory VectaMount™ Mounting Medium from Vecta Labs. Be sure to order your target C1 and C2 probes separately.
- RNAscope Intro Pack 2.5 Duplex Reagent Kit Human (cat. no 322435)
- RNAscope Intro Pack 2.5 Duplex Reagent Kit Mouse (cat. no 322436)
The RNAscope 2.5 HD Duplex Manual Assay Workflow Requires:
- RNAscope Target Probes (Catalog or Made-to-Order C1 & C2 Probes, compatitle with the same RNAscope probes used previously with RNAscope 2.0 2-plex assays.)
- RNAscope Duplex Positive Control Probes
- RNAscope Duplex Negative Control Probes (Cat. No. 320751)
- RNAscope Control Slides (optional)
- RNAscope 2.5 HD Duplex Reagent Kit (Cat. no 322430) (Includes Pretreatment Kit for FFPE samples, detection kit and wash buffer)
- Vector Labs Vectamount Permanent Mounting Medium (Cat. No. 321584)
- ImmEdge Hydrophobic Barrier Pen (Cat. No 310018)
- HybEZ™ Hybridization System
- Brightfield microscope
