Isolation of human B cell populations.

To study the function and biology of human B cells, it is necessary to isolate pure populations. Historically, B cells were enriched by the sequential depletion of monocytes, natural killer cells, and T cells. However, this time-consuming process has been superseded by negative selection methods using antibody cocktails targeted against other cell types or by positive selection using antibodies targeting B cell markers such as CD19. Here we detail three methods of isolating B cells from human blood or mononuclear cells and describe how these techniques can be combined with fluorescent cell sorting for the characterization of specific B cell populations.