Semi-quantitative Determination of Protein Expression Using Immunohistochemistry Staining and Analysis: An Integrated Protocol

Abstract

Semi-quantitative immunohistochemistry (IHC) is a powerful method for investigating protein expression and localization within tissues that involves using software, such as the freely available Fiji (ImageJ), to conduct deconvolution and downstream analysis. Currently, there is lack of an integrated protocol that includes a detailed procedure on how to measure or compare protein expression. Publications that use semi-quantitative methods to ascertain protein expression often don’t provide enough details in their methods section, which makes it difficult for the reader to reproduce their data. The current protocol provides an example and detailed steps of conducting semi-quantitative analysis of IHC images using Fiji software.