Fractionation of the non-histone proteins of antigen-reactive human lymphocytes: Criteria for the specificity of antigen uptake

Arye Rubinstein, John G. Gatien, Ezio Merler

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Abstract

Non-histone proteins were separated from the nuclei of lymphocytes for the purpose of testing if antigen, following antigen uptake, could be localized within this fraction. Human lymphocytes were disrupted by N2 cavitation, the nuclei isolated by differential centrifugation, the chromatin purified, and the DNA separated from chromatin proteins by chromatography on Bio-Gel A 5M in the presence of 4 M guanidine-HCl. Non-histone proteins were separated from the histones by chromatography on CM-Sephadex C-50 by a stepwise elution. Non-histone and histone proteins were obtained in satisfactory purity as shown by amino acid analysis and gel electrophoresis. Labeled antigen could be separated unaltered following its incubation with T-lymphocytes. On the other hand, antigen mostly degraded but still capable of precipitating with antibody was separated bound to the non-histone proteins of B-cells. Antigen was not bound to histones.

Original languageEnglish (US)
Pages (from-to)59-65
Number of pages7
JournalImmunochemistry
Volume13
Issue number1
DOIs
Publication statusPublished - Jan 1976

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ASJC Scopus subject areas

  • Medicine(all)

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