A new multiphasic buffer system for benzyldimethyl-n-hexadecylammonium chloride polyacrylamide gel electrophoresis of proteins providing efficient stacking.
Acidic PAGE systems using cationic detergents such as benzyldimethyl-n-hexadecylammonium chloride (16-BAC) or CTAB have proven useful for the detection of methoxy esters sensitive to alkaline pH, resolving basic proteins such as histones and membrane proteins. However, the interesting phosphate-based system suffered from poor stacking, resulting in broadened bands and long running times. Therefore, a new 16-BAC PAGE system based on the theory of moving boundary electrophoresis with properties comparable to the classical SDS-PAGE system was designed. As a result a new multiphasic analytical 16-BAC PAGE system providing efficient stacking and significantly shorter running times is presented here. It is based on acetic acid and methoxyacetic acid as common ion constituents. This PAGE system takes advantage of the additional counter stacking effect due to a cross boundary electrophoresis system resulting from the selected buffer constituents. Furthermore, the concentration of 16-BAC was optimized by determining its previously unknown CMC. Due to efficient focusing of the introduced tracking dye, methyl green, termination of electrophoresis can now be more easily followed as compared to the Schlieren line.
Kramer
Michael L
ML
Prion and Dementia Research Unit, University of Goettingen, Goettingen, Germany. mkramer@med.uni-goettingen.de
D000818:Animals; D001923:Brain Chemistry; D002021:Buffers; D004396:Coloring Agents / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005233:Fatty Alcohols; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D051844:alpha-Synuclein / Q000032:analysis
D002021:Buffers; D004396:Coloring Agents; D003902:Detergents; D005233:Fatty Alcohols; D011506:Proteins; D000644:Quaternary Ammonium Compounds; D051844:alpha-Synuclein; C012817:cetalkonium chloride