An alternative method of panning for rat B lymphocytes

  • Severson C
  • Burg D
  • Lafrenz D
 et al. 
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Abstract

A new method of panning for B lymphocytes is described in which the ability of the sIg+cells to adhere depends on the nature and concentration of nonspecific protein used rather than on the use of anti-immunoglobulin. Rat lymph node cells were suspended in 3% bovine serum albumin in Tris-buffered Hanks' and incubated in tissue culture flasks to allow adherence to the plastic. The recovered bed of adherent cells was shown by flow cytometry to be greater than 90% surface immunoglobulin positive and MHC class II positive while containing very few T cells. This adherent fraction was subsequently treated with anti-T cell antibody plus baby rabbit complement to produce a highly purified sIg+cell population containing no detectable T cells. The sIg+cells obtained by this panning procedure were functionally active in BCGF and BCDF assays. This method provides an easy and inexpensive alternative to conventional panning with anti-immunoglobulin and also eliminates the possibility of B cell activation by exposure to anti-immunoglobulin-coated surfaces. © 1987.

Author-supplied keywords

  • B cell
  • Differentiation
  • Panning
  • Purification
  • Rat

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Authors

  • C. D. Severson

  • D. L. Burg

  • D. E. Lafrenz

  • T. L. Feldbush

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