The RNAscope® Multiplex Assay uses a novel and proprietary method of in situ hybridization (ISH) to visualize single RNA molecules per cell in fresh frozen or embedded tissue samples. This assay provides exceptional sensitivity, allowing single-molecule detection of multiple RNA targets simultaneously and is ideal for co-localization studies of any expressed gene set in nearly any tissue type. The RNAscope® Multiplex Assay is developed by Advanced Cell Diagnostics and is distributed by Life Technologies.

  • Simultaneous detection of up to four RNA species
  • Single-molecule detection
  • Ready-to-use reagents
  • Compatible with fresh or embedded tissues

RNAscope® Multiplex Assay Kits

 

Each RNAscope® Multiplex Fluorescent Reagent Kit provides enough reagents to stain ~20 tissue sections approximately 20 mm x 20 mm large. Larger tissue sections will result in fewer tests. Select target-specific probes and housekeeping probes to analyze your genes of interest using the RNAscope® Multiplex Fluorescent Reagent Kit. Each kit contains:

  • Pretreatment Kit
  • Detection Kit
  • Wash Buffer Kit

  Our library of over 27,000 pre-designed target probes and made-to-order service ensure we can provide an assay for virtually any expressed gene.  Use the web form to request your probe set. We will typically respond within 24 hours with delivery details and terms.
 


Use the web form to request your probe set. We will typically respond within 24 hours with delivery details and terms.

Download Web Form

RNAscope® Multiplex Assay Applications

RNAScope® Multiplex Assays use common reagents and protocols to provide universal assay conditions for different target genes. Using our proprietary probe design, we provide double "ZZ" oligo probes designed to recognize your specific RNA target of interest. The images below show multiplexed results in gene expression analysis, neuroscience, target identification, toxicology, or tumor heterogeneity.

RNAscope® Multiplex Assays image gallery

Figure 1. PGK, PPIB, POLR2A, and HPRT1 mRNA, human breast cancer FFPE tissue. A multiplex fluorescent image from FFPE human breast cancer sections is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Phosphoglycerate Kinase (PGK), Peptidylprolyl isonerase B (PPIB), RNA Polymerase II DNA directed polypeptide A (POLR2A), and Hypozanthine phosphoribosyltransferase (HPRT1) mRNA expression. PGK mRNA is showed as green color; PPIB mRNA is showed in pink, PolR2A mRNA is showed in red, and PolR2A mRNA is showed in Yellow. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.

Figure 2. Beta-Actin, HPRT1, RPLP0, and PPIB expression in HeLa cell with RNAscope Multiplex Fluorescent Assay. A multiplex fluorescent image from Hela culture cell is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Large ribosomal protein (RPLPO), Beta-actin, Hypozanthine phosphoribosyltransferase (HPRT1), and Peptidylprolyl isonerase B (PPIB), and mRNA expression. RPLPO mRNA is showed as green color; Beta-actin mRNA is showed in red, HPRT1 mRNA is showed in yellow, and PPIB mRNA is showed in cyan. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.

Figure 3. IL-8 and beta-Actin mRNA expression in stimulated cultures cells with RNAscope Multiplex Fluorescent Assay. A multiplex fluorescent image from Hela culture cell is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Interleukin 8 (IL-8) and Beta-actin mRNA expression. IL-8 mRNA is showed as green color and Beta-actin mRNA is showed in red. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.

Figure 4. Fexf2 and NPY mRNA expression in mouse brain fresh frozen tissue with RNAscope Multiplex Fluorescent Assay. A multiplex fluorescent image from mouse fresh frozen brain tissue is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Fexf2 and Neuropeptide Y (NPY) mRNA expression.  Fexf2 mRNA is showed as green color and NPY mRNA is showed in red. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.


 

Figure 5. TH63 and SCNA mRNA expression in mouse cerebellum with RNAscope Multiplex Fluorescent Assay. A multiplex fluorescent image from mouse fresh frozen brain tissue is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Thyroid Hormone (TH63) and synuclein alpha (SCNA) mRNA expression.  TH63 mRNA is showed as green color and SCNA mRNA is showed in red. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.

Figure 6.  NPY, APOE, and Fezf2 mRNA expression in mouse brain fresh frozen tissue with RNAscope Multiplex Fluorescent Assay. A multiplex fluorescent image from mouse fresh frozen brain tissue is counterstained with DAPI. The tissue is stained with gene specific target probes to detect Neuropeptide Y (NPY), Apolipoprotein E (APOE), and FEZ family zinc finger 2 (Fezf2) mRNA expression. NPY mRNA is showed as green color, APOE mRNA is showed in red, and Fezf2 mRNA is showed in cyan. Image is monitor with 40X objective on Zeiss Axio imager and taken/composite with Nuance FX multispectral imaging system.

 Resources

 Molecular Probes® Handbook
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