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. 1979 Sep 1;181(3):759–761. doi: 10.1042/bj1810759

Dodecyl sulphate/polyacrylamide-gel electrophoresis at low pH values and low temperatures.

R Lichtner, H U Wolf
PMCID: PMC1161216  PMID: 42388

Abstract

A simple method is described for dodecyl sulphate/polyacrylamide-gel electrophoresis of pH- and temperature-labile biological intermediates. The method is based on a catalyst system that works at temperatures of 2--4 degrees C and pH values of 2--4 and an appropriate buffer system containing Li+ or Tris [CH2OH--C(CH2OH)2--NH3+] instead of Na+. This system does not lead to the precipitation of 1% dodecyl sulphate.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Avruch J., Fairbanks G. Demonstration of a phosphopeptide intermediate in the Mg ++ -dependent, Na + - and K + -stimulated adenosine triphosphatase reaction of the erythrocyte membrane. Proc Natl Acad Sci U S A. 1972 May;69(5):1216–1220. doi: 10.1073/pnas.69.5.1216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bader H., Sen A. K., Post R. L. Isolation and characterization of a phosphorylated intermediate in the (Na+ + K+) system-dependent ATPase. Biochim Biophys Acta. 1966 Apr 12;118(1):106–115. doi: 10.1016/s0926-6593(66)80149-2. [DOI] [PubMed] [Google Scholar]
  3. Choules G. L., Zimm B. H. An acrylamide gel soluble in scintillation fluids: its application to electrophoresis at neutral and low pH. Anal Biochem. 1965 Nov;13(2):336–344. doi: 10.1016/0003-2697(65)90202-2. [DOI] [PubMed] [Google Scholar]
  4. Drickamer L. K. The red cell membrane contains three different adenosine triphophatases. J Biol Chem. 1975 Mar 10;250(5):1952–1954. [PubMed] [Google Scholar]
  5. Fairbanks G., Avruch J. Four gel systems for electrophoretic fractionation of membrane proteins using ionic detergents. J Supramol Struct. 1972;1(1):66–75. doi: 10.1002/jss.400010110. [DOI] [PubMed] [Google Scholar]
  6. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  7. Hjertén S., Jerstedt S., Tiselius A. Electrophoretic "particle sieving" in polyacrylamide gels as applied to ribosomes. Anal Biochem. 1965 May;11(2):211–218. doi: 10.1016/0003-2697(65)90007-2. [DOI] [PubMed] [Google Scholar]
  8. Hokin L. E., Dahl J. L., Deupree J. D., Dioxon J. F., Hackney J. F., Perdue J. F. Studies on the characterization of the sodium-potassium transport adenosine triphosphatase. X. Purification of the enzyme from the rectal gland of Squalus acanthias. J Biol Chem. 1973 Apr 10;248(7):2593–2605. [PubMed] [Google Scholar]
  9. Jordan E. M., Raymond S. Gel electrophoresis: a new catalyst for acid systems. Anal Biochem. 1969 Feb;27(2):205–211. doi: 10.1016/0003-2697(69)90024-4. [DOI] [PubMed] [Google Scholar]
  10. Knauf P. A., Proverbio F., Hoffman J. F. Chemical characterization and pronase susceptibility of the Na:K pump-associated phosphoprotein of human red blood cells. J Gen Physiol. 1974 Mar;63(3):305–323. doi: 10.1085/jgp.63.3.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Narayan K. A., Vogel M., Lawrence J. M. Disk electrophoresis of wheat flour proteins with a modified apparatus utilizing gels of rectangular cross section. Anal Biochem. 1965 Sep;12(3):526–541. doi: 10.1016/0003-2697(65)90219-8. [DOI] [PubMed] [Google Scholar]
  12. Schuurmans Stekhoven F. M., van Heeswijk M. P., de Pont J. J., Bonting S. L. Studies on (Na+ + K+)-activated ATPase. XXXVIII. A 100 000 molecular weight protein as the low-energy phosphorylated intermediate of the enzyme. Biochim Biophys Acta. 1976 Jan 23;422(1):210–224. doi: 10.1016/0005-2744(76)90020-6. [DOI] [PubMed] [Google Scholar]
  13. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  14. Wolf H. U., Dieckvoss G., Lichtner R. Purification and properties of high-affinity Ca2+-ATPase of human erythrocyte membranes. Acta Biol Med Ger. 1977;36(5-6):847–858. [PubMed] [Google Scholar]
  15. Wolf H. U. Studies on a Ca 2+ -dependent ATPase of human erythrocyte membranes. Effects of Ca 2+ and H + . Biochim Biophys Acta. 1972 May 9;266(2):361–375. doi: 10.1016/0005-2736(72)90094-6. [DOI] [PubMed] [Google Scholar]

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