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The two‐dimensional pattern of cellular proteins from mitogen stimulated human peripheral blood lymphocytes
Author(s) -
Waldinger Dorothea,
Braun Andreas,
Cleve Hartwig
Publication year - 1985
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150061207
Subject(s) - silver stain , pokeweed mitogen , staining , biology , peripheral blood , stimulation , microbiology and biotechnology , mitogen activated protein kinase , lymphocyte , electrophoresis , immunology , concanavalin a , biochemistry , endocrinology , in vitro , genetics
Human peripheral blood lymphocytes were isolated from healthy donors and cultivated in the absence and presence of phytohemagglutinin (PHA) or pokeweed mitogen (PWM). Cell lysates of unlabelled lymphocytes or following stimulation of [H]leucine metabolically labelled lymphocytes were subjected to two‐dimensional electrophoresis. Differences in the protein patterns are described which are related to the method applied for visualization of the proteins (silver staining or fluorography), to the culture conditions (unstimulated, PHA or PWM stimulated lymphocytes) and to individual variations. Comparative analysis of the resulting patterns on silver stained gels and fluorograms revealed three qualitative and several quantitative differences. Comparing the two‐dimensional patterns of unstimulated and mitogenstimulated lymphocytes with silver staining, we recognized the quantitative increase of 6 protein spots after stimulation. Whereas protein patterns of PHA and PWM stimulated cells showed only minor quantitative differences when compared in silver stained gels, one qualitative and two quantitative differences between the two mitogens could be observed in fluorograms. Individual variability was generally low: within the approximately 30 unrelated individuals examined, thus far, the presence of two polymorphic proteins was demonstrated.