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.
