Detection of cellulose-binding proteins in electrophoresis gels by filter paper affinity blotting.
Proteins separated in electrophoresis gels were tested for the ability to bind cellulose by a simple blotting procedure. Proteins were blotted onto Whatman No. 1 filter paper by diffusion or by electrophoretic transfer and detected by Coomassie blue staining. Certain proteins released into culture supernatant by Bacteroides succinogenes NR9 (ATCC 43854) adhered strongly to cellulose, but were not found to have carboxymethylcellulose activity. Boiling of samples prior to electrophoresis eliminated the ability of proteins to bind to cellulose. Proteins that did not adhere to filter paper cellulose were detected on a nitrocellulose membrane placed behind the filter paper during electrophoretic transfer. The technique, referred to as filter paper affinity blotting, detects cellulose-binding proteins with great sensitivity.
Montgomery
L
L
Department of Animal Sciences, University of Illinois, Urbana 61801.
Fu
Y K
YK
D001426:Bacterial Proteins / Q000302:isolation & purification; D001439:Bacteroides / Q000032:analysis; D015153:Blotting, Western / Q000379:methods*; D002480:Cellulase; D004591:Electrophoresis, Polyacrylamide Gel; D005374:Filtration / Q000295:instrumentation; D006026:Glycoside Hydrolases / Q000302:isolation & purification; D010209:Paper; D012394:Rosaniline Dyes
D001426:Bacterial Proteins; D012394:Rosaniline Dyes; C048139:Coomassie blue; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase