Analysis of protein incorporation of radioactive isotopes in the Chinese hamster ovary cell cycle by electronic sorting and gel microelectrophoresis

Cytometry
J L PipkinD A Casciano

Abstract

The patterns of [3H]-leucine and [32P]-phosphate incorporation of proteins extracted with varying molarities of sodium chloride were analyzed from nuclei physically sorted from six fluorescence windows after propidium iodine staining of the G0 + G1 and G2 + M phases of the Chinese hamster ovary (CHO) cell cycle. Eight hundred nanograms of protein were used in each electrophoretic analysis obtained from 200,000 nuclei, a portion of the sample, from each window. Autoradiography was performed in a two-dimensional polyacrylamide gel ultra-microelectrophoresis apparatus (UMEA) designed and fabricated in this laboratory. There was a net reduction and/or loss of [3H]-leucine- and [32P]-phosphate-labeled protein regions from the autoradiographs occurring primarily in the G2 + M phase. Two phosphorylated proteins that were stage specific were observed in partitions of the G2 + M phase. The use of isolated proteins and the coelectrophoresis of these markers demonstrated the similarity in mobility of a number of proteins seen in the autoradiographs of proteins extracted with high and low salt molarities and implied they are synonymous. Coelectrophoresis indicated that a substantial number of high molecular weight proteins that decreased o...Continue Reading

References

Mar 1, 1976·Scientific American·L A HerzenbergL A Herzenberg
May 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·L R OrrickH Busch
Oct 15, 1973·Biochemical and Biophysical Research Communications·H Fujitani, V Holoubek
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Oct 1, 1969·Analytical Biochemistry·S BramhallJ R Loewenberg
Apr 1, 1984·Cell Biology International Reports·J L PipkinE R Burns

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Citations

Jun 1, 1987·Developmental Biology·J F AnsonD A Casciano
Jan 1, 1995·Life Sciences·J B LaBordeD K Hansen
Jan 1, 1988·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·J L PipkinD A Casciano
Oct 1, 1996·Aging : Clinical and Experimental Research·J L PipkinD A Casciano
Sep 1, 1992·Experimental Cell Research·J L PipkinD A Casciano
Jul 11, 2006·Folia Microbiologica·Y V KarpenkoN N Zhdanova
Aug 1, 1996·Aging : Clinical and Experimental Research·J L PipkinD A Casciano

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