Learn more about our New BaseScopeTM assay

Dual ISH and IHC

Human lung cancer with human PPIB positive control mRNA probe in red, CAM5.2 acidic cytokeratin immunohistochemical staining in green with DAPI as counterstain.

Human lung cancer with human PPIB positive control mRNA probe in red, CAM5.2 acidic cytokeratin immunohistochemical staining in green with DAPI as counterstain.

Dual ISH-IHC - Detection of OLFM4 using RNAscope technology and Lyzozyme using IHC

Dual ISH-IHC - Detection of OLFM4 using RNAscope technology and Lyzozyme using IHC

Dual ISH-IHC. Detection of alpha-smooth muscle actin (SMA) using IHC (green) and Olfr558 using RNAscope technology (red)

Dual ISH-IHC. Detection of alpha-smooth muscle actin (SMA) using IHC (green) and Olfr558 using RNAscope technology (red)

Analysis of both RNA and protein provides valuable information for understanding the regulation of expression. Both ISH and IHC technologies share common features that make them unique among the protein and RNA detection technologies.

    - Morphological context: detection of RNA and protein while conserving the tissue and cell structure

    - Similar workflow from sample fixation, pre-treatment, hybridization, signal amplification and data analysis.

Due to the multiple benefits of running both ISH and IHC, and the high robustness of RNAscope assay, some customers developed a dual ISH-IHC protocol to perform both analyses on the same sample slide. We conducted an interview with 4 researchers who developed their own dual ISH-IHC protocol to provide you some insides on their protocol.

Download the interview and learn from these experts how to develop a dual ISH-IHC protocol.

Interview with:

  • Kristie Wetzel, Novartis Institutes for

    BioMedical Research, Cambridge, MA, USA.

  • Dr Bradley Spencer-Dene, The Francis Crick

    Institute, London, UK.

  • Wanda Wang, University of Pittsburgh, PA,

    USA.

  • Mike Millar, Queen’s Medical Research

    Institute,University of Edinburgh,

    Edinburgh, UK.

Recorded Webinar

Application of combined RNA in situ hybridization and multiplex immunohistochemistry to unravel tumor heterogeneity in prostate cancerPresented by Dr Nallasivam Palanisamy and Dr Courtney Anderson

Watch Webinar

Publications using Dual ISH-IHC protocol

  1. Expression of olfactory signaling genes in the eye. Pronin et al.  Plos One. 2014 April 30;.9, e96435
  2. Hippo pathway activity influences liver cell fate. Yimlamai et al. Cell. 2014 Jun 5;157(6), 1324-1338 
  3. Simultaneous visualization and cell-specific confirmation of RNA and protein in the mouse retina. Stempel AJ. et al. Mol Vis. 2014 Sep 21;20: 1366-73
  4. Novel RNA hybridization method for the in situ detection of ETV1, ETV4, and ETV5 gene fusions in prostatecancer. Kunju LP. et al. Appl Immunohistochem Mol Morphol. 2014 Sep;22(8):e32-40
  5. Reporter mouse strain provides a novel look at angiotensin type-2 receptor distribution in the central nervous system. de Kloet AD. et al. Brain Struct. Funct. 2014 Nov 27
  6. A method for combining RNAscope in situ hybridization with immunohistochemistry in thick free-floating brain sections and primary neuronal cultures. Grabinski TM. et al. PLoS One. 2015 Mar20;10(3)
  7. The longitudinal transcriptomic response of the substantia nigra to intrastriatal 6-hydroxydopamine reveals significant upregulation of regeneration-associated genes. Kanaan NM. et al. PLoS One. 2015 May 20;10(5) 
  8. Interaction of the androgen receptor, ETV1, and PTEN pathways in mouse prostate varies with pathological stage and predicts cancer progression. Higgins, J.et al. Horm Cancer 2015 Jun;6(2-3), 67-68
  9. Semaphorin-3C signals through Neuropilin-1 and PlexinD1 receptors to inhibit pathological angiogenesis. Yang et al. EMBO Mol Med. 2015 Jul 20.
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