What is immunohistochemistry and in situ hybridization?
Immunohistochemistry is the detection of a protein of interest in thin tissue sections or cells mounted on slides for microscopic evaluation. In situ hybridization is also done on thin tissue sections or cells mounted on slides, but it detects a specific sequence or region of DNA or RNA.
What is the difference between IHC and ISH?
ISH is a nucleic acid hybridization technique which is directly performed on a portion or section of tissue or the entire tissue. IHC is a technique where monoclonal and polyclonal antibodies are utilized to determine the presence of antigens, which are special protein markers placed on the cell surfaces.
What would you use in situ hybridization for?
In situ hybridization is used to reveal the location of specific nucleic acid sequences on chromosomes or in tissues, a crucial step for understanding the organization, regulation, and function of genes.
What does IHC stain for?
Immunohistochemistry (IHC) is the most common application of immunostaining. It involves the process of selectively identifying antigens (proteins) in cells of a tissue section by exploiting the principle of antibodies binding specifically to antigens in biological tissues.
How long should an in situ probe be?
250–1,500 bases
RNA probes should be 250–1,500 bases in length; probes of approximately 800 bases long exhibit the highest sensitivity and specificity. Transcription templates should allow for transcription of both probe (antisense strand) and negative control (sense strand) RNAs.
What probes are used in in situ hybridization?
In situ hybridization probes
- Double-stranded DNA (dsDNA) probes.
- Single-stranded DNA (ssDNA) probes.
- RNA probes (riboprobes)
- Synthetic oligonucleotides (PNA, LNA)
What does Northern blot tell you?
A northern blot is a laboratory method used to detect specific RNA molecules among a mixture of RNA. Northern blotting can be used to analyze a sample of RNA from a particular tissue or cell type in order to measure the RNA expression of particular genes.
Why is IHC used?
Immunohistochemistry (IHC) is an important application of monoclonal as well as polyclonal antibodies to determine the tissue distribution of an antigen of interest in health and disease. IHC is widely used for diagnosis of cancers; specific tumor antigens are expressed de novo or up-regulated in certain cancers.