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Bio-Rad Immunophenotyping of B Cells

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Immunophenotyping of B Cells by Flow Cytometry

Common markers and gating strategies

B lymphocytes along with T lymphocytes form the adaptive immune system. They are a heterogeneous population of cells with unique functional properties. Their primary function is to produce antibodies, however they can also be professional antigen presenting cells to T lymphocytes helping to generate an effective immune response. Abnormal B cell development or function can result in several autoimmune diseases and malignancies, and their identification can be critical for disease treatment and detection of residual disease.

B cell subsets can be difficult to identify due to variability and low level expression of markers and the rarity of some populations. This guide to immunophenotyping of human B cells takes you through some of the common markers and gating strategies used to identify B cells by flow cytometry, with examples of data acquired on the ZE5™ Cell Analyzer.


The first marker to be identified, using monoclonal antibodies specific for B cells, was B1, now known as CD20. However the key marker for identification of human B cells is CD19, which is expressed by virtually all B cell lineages (in mouse the key pan B cell marker is CD45R, also known as B220, and in rat it is CD45RA). With the addition of more markers, such as CD27, CD24, CD38 and surface immunoglobulin, specific blood subsets such as naïve, memory and transitional B cells can be identified.

Additional markers allow identification of secondary lymphoid organ B cell populations, for instance CD24 and CD21, for marginal zone B cells, and CD69, CD80 and CD86 for activation status. For more information on B cell lineage markers, function and activation, including all our antibodies useful for identification of B cells please go to our dedicated B cell page.

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