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. 1983 Jan;80(2):524–528. doi: 10.1073/pnas.80.2.524

Stable transformation of mouse L cells for human membrane T-cell differentiation antigens, HLA and beta 2-microglobulin: selection by fluorescence-activated cell sorting.

P Kavathas, L A Herzenberg
PMCID: PMC393411  PMID: 6188154

Abstract

We isolated stable transformants of mouse L cells expressing human cell surface differentiation antigens by using immunofluorescence with monoclonal antibodies and selection with a fluorescence-activated cell sorter (FACS). Mouse L cells (TK-) were cotransformed with human cellular DNA and the herpes simplex virus thymidine kinase (TK) gene. TK+ transformants were first selected. The TK+ populations were stained with various fluorescent antibodies to membrane antigens, and positive cells were sorted and cloned by using a FACS. Transformants for HLA class I antigens, for beta 2-microglobulin, and for the T-cell differentiation antigens Leu-1 and Leu-2 were isolated. The frequency of antigen transformants among the TK+ transformants was about 0.5 X 10(-3). The sizes of the HLA, Leu-1, and Leu-2 molecules expressed by the transformants were the same as those of the proteins present on DNA-donor cells.

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Selected References

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