BAC-PAGE

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The BAC-PAGE is a biochemical method for the separation of proteins according to their molecular weight in a polyacrylamide gel electrophoresis (PAGE) using the surfactant 16-benzyldimethylhexadecyl (16-BAC).

properties

The BAC-PAGE is suitable for the separation of proteins with many positive charges or glycosylated membrane proteins and for 2D gel electrophoresis of membrane proteins, including the separation of mitochondrial membrane proteins. In contrast to the commonly used SDS-PAGE , the proteins loaded with the cationic surfactant 16-BAC migrate to the cathode (minus pole) and the gel buffer and the electrophoresis buffer have an acidic pH . Alternatively, CTAB-PAGE can be used instead of BAC-PAGE as PAGE with a cationic surfactant . If the samples are not heated with the sample buffer and no reducing agent such as dithiothreitol or dithioerythritol has been added, the native conformation and biological activity of the proteins is partially retained.

Individual evidence

  1. ↑ DE Macfarlane: Use of benzyldimethyl-n-hexadecylammonium chloride ("16-BAC"), a cationic detergent, in an acidic polyacrylamide gel electrophoresis system to detect base labile protein methylation in intact cells. In: Analytical biochemistry. Volume 132, Number 2, July 1983, pp. 231-235, PMID 6625164 .
  2. ↑ Joachim Hartinger, Katinka Stenius, Dagmar Högemann, Reinhard Jahn : 16-BAC / SDS-PAGE: a two-dimensional gel electrophoresis system suitable for the separation of integral membrane proteins. In: Analytical Biochemistry. Vol. 240, No. 1, 1996, pp. 126-133, PMID 8811889 , doi : 10.1006 / abio.1996.0339 .
  3. ^ RP Zahedi, J. Moebius, A. Sickmann: Two-dimensional BAC / SDS-PAGE for membrane proteomics. In: Sub-cellular biochemistry. Volume 43, 2007, pp. 13-20, PMID 17953388 .
  4. ↑ RJ Brown, N. Kinkl, M. Beer, M. Ueffing: Two-dimensional electrophoresis of membrane proteins. In: Analytical and bioanalytical chemistry. Volume 389, Number 4, October 2007, pp. 1033-1045, doi : 10.1007 / s00216-007-1514-6 , PMID 17680235 .
  5. ↑ HG Nothwang, J. Schindler: Two-dimensional separation of membrane proteins by 16-BAC-SDS-PAGE. In: Methods in molecular biology (Clifton, NJ). Volume 528, 2009, pp. 269-277, doi : 10.1007 / 978-1-60327-310-7_19 , PMID 19153699 .
  6. ↑ B. Wenge, H. Bönisch, J. Grabitzki, G. Lochnit, B. Schmitz, MH Ahrend: Separation of membrane proteins by two-dimensional electrophoresis using cationic rehydrated strips. In: Electrophoresis. Volume 29, Number 7, April 2008, pp. 1511-1517, doi : 10.1002 / elps.200700546 , PMID 18318447 .
  7. ↑ RJ Brown, N. Kinkl, H. Zischka, M. Ueffing: 16-BAC / SDS-PAGE analysis of membrane proteins of yeast mitochondria purified by free flow electrophoresis. In: Methods in molecular biology (Clifton, NJ). Volume 528, 2009, pp. 83-107, doi : 10.1007 / 978-1-60327-310-7_6 , PMID 19153686 .
  8. ↑ Engelbert Buxbaum: Cationic electrophoresis and electrotransfer of membrane glycoproteins. In: Analytical Biochemistry. Vol. 314, No. 1, 2003, pp. 70-76, PMID 12633604 , doi : 10.1016 / S0003-2697 (02) 00639-5 .
  9. ↑ Dianne T. Akin, Raymond Shapira, Joseph M. Kinkade Jr .: The determination of molecular weights of biologically active proteins by cetyltrimethylammonium bromide-polyacrylamide gel electrophoresis. In: Analytical Biochemistry. Vol. 145, No. 1, 1985, pp. 170-176, PMID 4003759 , doi : 10.1016 / 0003-2697 (85) 90343-4 .
  10. ^ RJ Simpson: CTAB-PAGE. In: Cold Spring Harbor protocols. Volume 2010, number 4, April 2010, p. Pdb.prot5412, doi : 10.1101 / pdb.prot5412 , PMID 20360366 .