Synthetic Antigen Gels as Practical Controls for Standardized andQuantitative Immunohistochemistry

摘要

Optimization and standardization of immunohistochemistry (IHC) protocols withinand between laboratories requires reproducible positive and negative controlsamples. In many situations, suitable tissue or cell line controls are notavailable. We demonstrate here a method to incorporate target antigens intosynthetic protein gels that can serve as IHC controls. The method can usepeptides, protein domains, or whole proteins as antigens, and is compatible witha variety of fixation protocols. The resulting gels can be used to create tissuemicroarrays (TMAs) with a range of antigen concentrations that can be used toobjectively quantify and calibrate chromogenic, fluorescent, or massspectrometry-based IHC protocols. The method offers an opportunity toobjectively quantify IHC staining results, and to optimize and standardize IHCprotocols within and between laboratories.