Special considerations for glycolipids and their purification.
This unit describes the antigenic stimulation of in vitro antibody production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro antibody production is presented along with a procedure for quantifying the number of antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.
Smith
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DF
The University of Georgia, Atlanta, Georgia, USA.
Prieto
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PA
D000818:Animals; D002477:Cells / Q000378:metabolism; D002853:Chromatography, Liquid; D006017:Glycolipids / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D013997:Time Factors
D006017:Glycolipids