Primary and Secondary Antibodies and Flow Cytometry Controls

Flow cytometry requires robust, sensitive, and specific monoclonal antibodies conjugated to bright, stable fluorochromes. In this chapter, we’ll discuss the development process to generate antibodies specifically geared for use in flow cytometry. We will delve into how this technique is used for detecting multiparameter surface marker expression on cells by outlining how to use unconjugated antibodies in flow. We will finish by describing the various controls that can be used for evaluating protein expression including isotype controls, lineage controls, activation controls, and fluorescence minus one.