The RNAscope® 2.5 LS Duplex Assay is designed as an automated assay on the Leica Biosystems' BOND RX Research Advanced Staining System. The RNAscope® 2.5 LS Duplex Assay, is an RNA in situ hybridization assay 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 a multitude of sample types mounted on slides. RNAscope® Assays do not require the RNA-free environment used for traditional ISH.

RNAscope® 2.5 LS Duplex Kit is designed for simultaneous in situ detection of two RNA species.

Common applications include co-localization studies to map co-expression of two targets within the same cellular context (e.g. secreted ligand and its receptor) or to profile gene expression in a specific cell type expressing a known marker (e.g. a specific stem cell marker). To distinguish between the two targets, individual probes are used in probe channels 1 (C1) and Channel 2 (C2). Channel 1 (C1)  is detected using HRP based diaminobenzidine (DAB) reaction resulting in a dark brown signal and Channel 2 (C2)  is detected using an AP Fast Red based reaction resulting in a red color. Due to the brightness of the red chromogen, it is advised that the lower expressed target be ordered in the C2 channel and stronger expressed target be ordered in the C1 channel.The stained slides are visualized with a bright-field microscope.

As an alternative to diaminobenzidine (DAB), one can also substitute the HRP-based green chromogen using the RNAscope® 2.5 LS Green Accessory Pack to result in a detectable green and red signal respectively.

These new RNAscope® 2.5 LS Duplex reagents require the new RNAscope® 2.5 LS Probes for optimal performance. Probe designs are identical to previous version, however the RNAscope® 2.5 LS probes are manufactured for optimal performance with RNAscope® 2.5 LS reagent chemistry.

Sample Images




RNAscope® 2.5 LS Duplex Reagent Kit Workflow Requires the following from ACD:

RNAscope® 2.5 LS Target Probes (Catalog or Made-to-Order LS Probes)
300038 RNAscope® 2.5 LS Probe (RTU)
300038-C2 RNAscope® 2.5 LS Probe- C2 (50x)
RNAscope® 2.5 LS Control Probes choose from Pooled Control Probes for both C1 and C2 channels or Control probes in C2 only:
​​​Pooled Control Probes
320768 RNAscope® 2.5 LS Duplex Control Probes (PPIB-C1, Polr2A-C2)- Mm

320748 RNAscope® 2.5 LS Duplex Control Probes (PPIB-C1, Polr2A-C2)- Hu

320758 RNAscope® 2.5 LS Duplex Negative Control Probe (DapB-C1, DapB-C2)

Control Probes for C2 only (to be used with C1 RTU Target probes)
313908-C2 RNAscope® 2.5 LS Positive Control Probe - Hs-PPIB-C2 (50X)

312038-C2 RNAscope® 2.5 LS Negative Control Probe - DapB-C2 (50x)

RNAscope® Control Slides (optional)
Human Cell Pellet Cat. No. 310045, or Mouse 3T3 Cell Pellet Cat. No. 310023
RNAscope® 2.5 LS Duplex Reagent Kit Cat No. 322440
RNAscope® 2.5 LS Green Accessory Pack Cat No. 322550

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.