Immunohistochemistry as an Important Technique in Experimental and Clinical Practices
- Authors: Hareram Birla, Sachchida Nand Rai, Saumitra Sen Singh, Walia Zahra, Neeraj Tiwari, Aijaz A. Naik, Anamika Misra, Shikha Bharati, Surya Pratap Singh9
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View Affiliations Hide Affiliations9 Department of Biochemistry, Institute of Science, Banaras Hindu University, Varanasi 221005, India
- Source: Protocols used in Molecular Biology , pp 44-59
- Publication Date: January 2020
- Language: English
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Immunohistochemistry (IHC) is a well-known technique in the field of biological and medical sciences. This technique is based on the principle of antigenantibody interaction and is used for identification of cellular or tissue constituents, i.e., an antigen by using a specific antibody. The binding of an antibody to an antigen is confirmed either by labelled primary antibody itself or by using secondary labelling method such as fluorescence labelled antibody. Such interactions give information about the cellular process occurring inside the cell. In last few years, huge amount of data have been generated using IHC. Furthermore, adequate knowledge of this technique is required for the optimum result and its reproducibility. The detailed information about the tissue section, antigen retrieval (AR), increased sensitivity of the detection systems and proper standardization are the key points for this technique. This protocol will address overview of the technique, tissue preparation, microtome, antigen retrieval, antibodies and antigen fixation, detection methods, background reduction and trouble shootings.
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