Wanja Kildal

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Immunohistochemistry (IHC)

Studying protein expression and localization of proteins in tissues.

One of the main goals for the ICGI is to examine the role of large scale genomic instability in cancer development. We describe large scale instability mainly through DNA ploidy and nucleotyping. From the literature it is well known that errors in the cell cycle, often through deregulation of mitosis and cytokinesis, can lead to DNA aneuploidy and genetic instability. To gain further information on the relationship between cell cycle regulators and aneuploidy, and to examine possible prognostic markers for patients with cancer, we conduct immunohistochemical (IHC) studies of selected proteins involved in for example cell cycle regulation on different tumor materials.

Method

By IHC one employs specific antibodies that bind to specific antigens to study the protein expression and localization of proteins in tissues. The method is widely used in diagnostic pathology. Different methods can be used for the visualization of the bound antibody. In the direct method a primary antibody conjugated to a reagent, such as an enzyme that can catalyze a color producing reaction or a fluorescent tag, binds to the antigen. In order to amplify the signal, the indirect visualization system uses a conjugated secondary antibody, which binds to an unlabeled primary antibody. Biotinylated secondary antibodies coupled with streptavidin/peroxidase are commonly used (Figure 1). The enzyme reacts with 3, 3’-Diaminobenzidine to produce a brown staining when the targeted antigen is present.   

Seeking Prognostic Biomarkers

We are currently using AutostainerLink 48 (Dako, Agilent technologies) for our IHC projects. We aim at identifying a large number of potential cancer related proteins in sections from patients with prostate and colorectal carcinomas. The relation between the protein expression and clinicopathological variables including survival for the patients will be examined, with a main goal of identifying potential prognostic bio­markers for patients with these cancer types.

This text was last modified: 16.03.2017

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Chief Editor: Prof. Håvard E. Danielsen
Copyright Oslo University Hospital. Visiting address: The Norwegian Radium Hospital, Ullernchausséen 64, Oslo. Tel: 22 78 23 20