Metastasis Research Protocols

Volume 878 of the series Methods in Molecular Biology pp 137-147


Determination of Cell-Specific Receptor Binding Using a Combination of Immunohistochemistry and In Vitro Autoradiography: Relevance to Therapeutic Receptor Targeting in Cancer

  • Michael R. DashwoodAffiliated withDepartment of Clinical Biochemistry, Royal Free and UCL Medical School Email author 
  • , Marilena LoizidouAffiliated withDivision of Surgery & Interventional Science, Royal Free and UCL Medical School

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Mapping of receptor binding to specific structures, or cells within tissue samples, provides valuable information regarding biological and pathological mechanisms. Such information may potentially be translated into targeted therapies, especially in the field of cancer treatment. In this chapter, a receptor localization technique is described which utilises frozen sections of human tissue and combines immunohistochemistry (IHC) and micro-autoradiography. IHC utilises antibodies tagged to an enzymatic complex to identify specific cell types (such as epithelial cells or fibroblasts) within the tissue under investigation; this step is immediately followed by the second technique which is based on the use of radiolabelled compounds (radioligands) that selectively bind to preselected membrane receptors. This approach allows visualisation of cells of interest by immunohistochemical staining of tissue sections (colour product) in combination with the use of radiolabelled compounds that are detected following exposure to radiation-sensitive film or emulsion to produce a map of receptor distribution or localisation of cell-specific receptor binding. The system described has been used to compare receptor binding to cells in normal human colorectal tissue with that in colorectal cancer specimens.

Key words

Cell type-specific immunohistochemistry Receptor micro-autoradiography Receptor binding