Immunohistochemistry (IHC) is a staining technique that utilizes specific antibodies to identify the presence or absence of specific protein antigens, using a chromogenic detection system. in situ hybridization (ISH) is a technique that uses probes that bind to target RNA or DNA sequences, detected by fluorescent or chromogenic dyes.
IHC and ISH are essential techniques that can help you understand the biology in your system at the tissue level.
IHC and/or ISH can help you to:
- Visualize the gene and/or protein expression in different cell types
- Visualize the effects of your morpholino or exon-skipping drug on tissue gene expression.
- Determine if cellular protein or RNA biomarkers track with disease severity or response to therapy
It isn’t enough to just perform a stain. It is very important to know what your antibody is staining. Research projects can get off track with spurious immunohistochemistry results.
Key Considerations for antibody or RNA-probe validation:
- Have you selected appropriate positive and negative tissue controls for your validation?
- Is your antibody staining the intended protein?
- Is the differential staining between target and non-target reproducible?
ALK-11 immunostain highlighting ALK overexpression in the nuclear membrane of an inflammatory myofibroblastic tumor with a RANBP2-ALK gene fusion. RANBP2 is a nuclear membrane protein.
Dr. Hill has extensive experience in immunohistochemistry. She was the first to automate the immunohistochemistry laboratory at St. Jude Children’s Research Hospital. At Resourcepath we use the Ventana Discovery Ultra platform for automated immunohistochemistry and in situ hybridization which makes it easy to transition validated antibody testing to other automated platforms.