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.

Sample Images




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

Bio-Techne Corporation (NASDAQ: TECH) is a leading developer and manufacturer of high quality purified proteins:notably cytokines and growth factors, antibodies, immunoassays, as well as biologically active small molecule compounds and ACD's in situ hybridization detection products,which are sold to biomedical researchers and clinical research laboratories; these operations constitute the core Biotechnology Division, headquartered in Minneapolis, Minnesota. The Protein Platforms Division manufactures innovative protein analysis tools under the ProteinSimple brand name that greatly automate western blotting and immunoassay practices. The Diagnostics Division manufactures FDA-regulated controls, calibrators, blood gas and clinical chemistry controls and other reagents for OEM customer and clinical customers. Bio-Techne products are integral components of scientific investigations into biological processes and the nature and progress of specific diseases. They aid in drug discovery efforts and provide the means for accurate clinical tests and diagnoses. With thousands of products in its portfolio, Bio-Techne generated approximately $563 million in net sales in fiscal 2017 and has approximately 1,800 employees worldwide. For more information on Bio-Techne and its brands, please visit www.bio-techne.com.