Purification and Characterization of Antigen-Binding Virgin and Memory B Cells

C. D. Myers, V. M. Sanders, E. Yefenof, K. G. Oliver, J. W. Uhr, E. S. Vitetta

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

This chapter focuses on the methods for purifying resting antigen-specific B cells so that their activation by antigens and T cells can be assessed. Many methods for studying the physiology and activation of B lymphocytes are available. The chapter describes a method for purifying a population of TNP-specific B cells (TNP-ABC) from spleens of normal or primed mice. These cells are suitable for the analysis of early activation events of B cells in response to antigen. However, to interpret such data, an accurate measurement of the purity of the enriched population is required. The antigen-binding capacity of these cells as measured by rosetting or binding fluorescent antigen gives a far higher estimate of purity than functional assays, such as proliferation, plaque formation, and an enumeration of clones in the splenic focus assay.

Original languageEnglish (US)
Pages (from-to)316-325
Number of pages10
JournalMethods in Enzymology
Volume150
Issue numberC
DOIs
StatePublished - Jan 1987

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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