Flow cytometry is one of the powerful tools for the investigators in immunological research involved in studying various immune cells. One of the main advantages of this technique is that is capable of multi-parameter measurements that can be accomplished on a single-cell basis. As a result of the advances in the flow cytometry researchers can now decrypt the phenotypes of several cell subsets in ways that were not possible using traditional assays, such as Western/immunoblotting, microarrays and enzyme-linked immunosorbent assays. Abundance of reagents and applications has led to a large array of ways in which flow cytometry is being used to screen and employ immunophenotyping assays to identify lymphocyte, monocyte, granulocyte and peripheral blood mononuclear cell subsets to name a few in humans and other model organisms. The choice of antibody cocktails and surface markers expressed on the cells is important in immunophenotyping. Use of different antibody clones and fluorochromes in immunophenotyping can greatly influence results, for example use of two different antibody clones from different sources might result disparate staining results for the same cells from a donor. It has been reported that fluorescence separation of positive and negative populations for a marker could vary depending on the flurochromes used as this accounts for the difference among the cells positive for a marker separated from a negative cohort of cells (1). Needless to say, the antibody titer, reagents used, sample handling and instrument setup also contributes to the variability in interpretation of the experimental data.
1. PMID: 15536642.
Novus Biologicals in the forefront in specializing and delivering exceptional immunophenotyping reagents and offer you the latest range of flurochromes along with antibodies and accessory buffers for your research needs.
