At the Human Protein Atlas project’s Rudbeck Laboratory, the Thermo Scientific Autostainer 480 has improved quality and reproducibility while increasing reliability and throughput, as Peter Brooks and Kenneth Wester explain.
Established to allow for systematic exploration of the human proteome using antibody-based proteomics, the Human Protein Atlas (HPA) project is hosted by the Swedish Human Proteome Resource (HPR) programme to perform enzyme-based immunohistochemical analysis. The HPA is a publicly available database with millions of high-resolution images showing the spatial distribution of proteins in 48 different normal tissues, 20 different cancer types and 47 different human cell lines.
The project was launched in 2005, and has since grown from 718 antibodies and 413,568 images to more than 8800 antibodies and 7,300,000 images. Committed to producing specific antibodies to human target proteins, the project provides scientists with a thorough insight into the function of different proteins and how changes in their distribution could be reflected in diseases such as cancer.
The project performs application-specific validation for each antibody using a variety of analytical techniques, including immunohistochemistry. The technique is implemented for large-scale protein profiling in tissues and cells at the Rudbeck immunohistochemistry laboratory of the Uppsala University in Sweden. Since 2007, the laboratory has evaluated approximately 700 new antibodies each month. Around half of these antibodies are judged unsuitable for immunohistochemistry application and as a result are not published on the Human Protein Atlas.
Immunohistochemistry
Immunostaining involves the use of antibodies for staining tissue samples to detect specific proteins. Immunohistochemistry (IHC) on tissue sections is the most commonly applied immunostaining technique and can be used for a variety of applications based on investigating the presence or absence of a protein, its tissue distribution, its subcellular localisation and/or changes in protein expression or degradation. The purpose of immunostaining is to enable the in situ detection of antigens in tissue sections and cells by lectins, monospecific monoclonal and polyclonal antibodies. Detection is obtained by visualisation of the antigen by a light microscope-detectable opaque dye.
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