Optimized Human on Human Immunodetection

Immunohistochemistry (IHC) remains a mainstay application for investigators in biological sciences. For the generation of reliable and reproducible data in IHC however, assay specificity and sensitivity are paramount. As investigators continue to explore antigen expression using IHC methodologies in a myriad of normal and disease animal models, xenograft systems and other projects involving different or same species combinations of antibodies and tissue specimens, use of appropriate controls and accurate reporting of staining results, without ambiguity, are vital.

Study Overview

The hapten tagged antibody approach is the predominant methodology currently in use for detecting human primary antibodies on human tissue sections for therapeutic antibody assessment.

This study utilizes frozen tissue sections to compare workflows and staining results between the hapten tagged method and the method referenced in the H.O.H.™ (Human on Human) Immunodetection Kit (HOH-3000) available from Vector Laboratories. A brief outline of the main reagents used to conduct the IHC assays comparing these two methods is also included. Unless otherwise noted, all reagents were from Vector Laboratories.

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Serial sections of human kidney (FFPE) showing strong, specific staining using human anti-cytokeratin primary antibody detected with HOH-3000 kit from Vector Laboratories
LEFT image: Serial sections of human kidney (FFPE) showing strong, specific staining using human anti-cytokeratin primary antibody detected with HOH-3000 (brown regions). RIGHT image: Negative control showing an absence of staining (no background).