Getting started in immunohistochemistry

This month at CiteAb we’re focusing on immunohistochemistry, and as this time of year sees a new intake of postgraduate researchers we thought we’d start at the beginning, with a small overview of the history of the discipline and details of some of the services we’ve personally found useful.

Immunohistochemistry, often referred to as IHC, is a progression from histochemical analysis relying on highly specific antibody-antigen interactions as a method of protein detection. This allows visualization of the distribution and localization of specific cellular components within a cell or tissue.

Histochemistry was widely in use for some time before the use of antibodies was introduced in 1939 by Dr Albert Coons. He came up with the idea of using antibodies to identify antigens during what he termed as a ‘scientific epiphany’ while on holiday in Berlin.

Dr Coons formalised his ideas in the paper ‘Immunological properties of an antibody containing a fluorescent group [2]’, followed by a second paper titled ‘The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody’. His work was then interrupted when he was drafted to the US Medical Corps during World War II.

Since it’s inception, IHC has seen an almost constant effort to improve sensitivity of detection and one of the major developments remains the introduction of indirect methods of staining, using peroxidase anti-peroxidase (PAP) in 1979 [4] and avidin-biotin complexes in 1981 [5].

Here at CiteAb we have used a number of resources which we think are particularly valuable. Dako has a great resource covering just about everything you might need to know of the theory of IHC right through to the fine details. There is also a good overview of immunohistochemistry on the Thermo Scientific website.

There are also a number of fluorescence spectra viewers that we’d recommend trying. Invitrogen provides a clean and easy to use spectra viewer with a great deal of choice of fluorophores, lasers and filters available for analysis. Other companies, including BD Bioscience and eBioscience, also have spectra viewers available, but as these require Java which doesn’t support the 32bit Chrome web browser we found Invitrogens offering to be the easier option.

The Arizona University Spectra Database is certainly one of the most comprehensive spectra viewers available, although the interface isn’t as pleasing as some of the other options.

Of course, you can use CiteAb to source antibodies. We actually have around 15,000 citations referencing the use of antibodies in various IHC methods, including frozen, paraffin, wholemount and more.

We’ll continue to explore immunohistochemistry in the weeks to come, so do keep coming back to our blog to see our latest updates. We’d also love for you to join in the conversation either in the comments here or on Twitter, using our handle @CiteAb. Let us know what resources you’ve found useful!

– the CiteAb team

 


References

1 – The National Academies Press. Biographical Memoirs. [online] Available at: <http://www.nap.edu/readingroom.php?book=biomems&page=acoons.html>[Accessed 28 August 2013]

2 – Coons AH, Creech HJ, Jones RN. Immunological properties of an antibody containing a fluorescent group. Proc Soc Exp Biol Med. 1941;47:200–202

3 – Coons AH, Creech HJ, Jones RN, Berliner E. The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody. J Immunol. 1942;45:159–170

4 – Nakene PK. Simultaneous localization of multiple tissue antigens using the peroxidase labeled antibody method: a study of pituitary glands of the rat. J Histochem Cytochem 1968;16:557-60

5 – Hsu SM, Raine L, and Fanger H. Use of avidin-biotin peroxidase complex (aBC) in immunoperoxidase techniques: a comparison between aBC and unlabeled antibody (PaP) procedures. J Histochem Cytochem 1981;29:577-80.
PMID: 6166661

6 – Albert H. Coons. The beginnings of immunofluorescence. J Immunol. 1961;87:499-503
PMID: 13881115

7 – Coons AH. The development of immunohistochemistry. Ann N Y Acad Sci. 1971 Jun 21;177:5-9.
PMID: 4400556


Subscribe to the CiteAb Newsletter