Dual ISH-IHC Antibody Validation

Bio-Techne continues to undertake rigorous steps to validate antibodies and ensure antibody reproducibility. By combining tissue staining strategies from the Advanced Cell Diagnostics™, R&D Systems™ and Novus Biologicals™ brands, we are taking a multiomics approach to antibody validation.

CD68/SR-D1 in Human Tonsil Using Dual RNAscope®ISH and IHC. CD68/SR-D1 mRNA (red) and protein (green) was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope® Probe (Catalog # 560591) and immunohistochemistry using R&D Systems Mouse Anti-Human CD68/SR-D1 Monoclonal Antibody (Catalog# MAB2040)

CD68/SR-D1 antibody in human tonsil using dual RNAscope ISH and IHC

Get More Data from a Single Slide Using Dual RNAscope™ ISH Technology with Antibodies for IHC

ISH-IHC Benefits

  • Dual RNAscopeTM ISH-IHC Co-Detection Assays virtually eliminate lengthy protocol optimizations and allow detecting proteins and RNA on the same slide
  • Conserve precious samples
  • Improves antibody performance allowing use of any immunohistochemistry (IHC) antibody
  • The specificity of protein detection in tissues by IHC antibodies can be confirmed by detecting RNA transcripts in tissue using RNAscope™ in situ hybridization (ISH).
  • Morphological context: detection of RNA and protein while preserving the cell and tissue structural organization
  • Antibody-independent validation, or "Orthogonal Validation", as proposed by the International Working Group for Antibody Validation (IWGAV) and the five pillars of antibody validation (Uhlen, M. et al. (2016) Nat. Methods 13:823)
  • Growing number of publications utilizing multiomics approach to investigating the regulation of gene expression using Dual RNAscope ISH-IHC.
Learn More About the Co-Detection Assays

ISH-IHC Co-detection Workflow

IHC-ISH workflow for sample preparation to analysis

Additional Dual ISH-IHC Resources: