Anaerobic multiphasic gel electrophoresis of the molybdoproteins in extracts of Clostridium pasteurianum.
Simple procedures using multiphasic buffer systems for anaerobic electrophoresis have been devised to identify oxygen-labile metalloproteins. An anaerobic slab gel apparatus was developed with cooling and design for anaerobic conditions. Included is a procedure to remove sample wells after stacking proteins in a crude extract, to prevent streaking (background) caused by continuous leakage of "nonstacked protein" from the sample wells. Identification of eleven Mo zones in extracts of Clostridium pasteurianum demonstrates the usefulness of the technique in identifying radiolabeled oxygen-labile proteins in cell-free crude lysates.
D000693:Anaerobiosis; D001345:Autoradiography; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002021:Buffers; D055598:Chemical Phenomena; D002621:Chemistry; D003013:Clostridium / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods; D008667:Metalloproteins / Q000302:isolation & purification*; D008982:Molybdenum / Q000032:analysis*
D001426:Bacterial Proteins; D002021:Buffers; D008667:Metalloproteins; D008982:Molybdenum