Sensitive detection of endo-1,4-beta-glucanases and endo-1,4-beta-xylanases in gels.
A simple, highly sensitive zymogram technique for detection of endo-1,4-beta-glucanases and endo-1,4-beta-xylanases in polyacrylamide gels after electrophoresis or isoelectric focusing was developed. The detection employs transparent agar replicas containing soluble covalently dyed polysaccharides, hydroxyethylcellulose dyed with Ostazin brilliant red H-3B and beechwood 4-O-methyl-D-glucurono-D-xylan dyed with Remazol brilliant blue R, as the respective substrates. The high sensitivity of the detection is achieved by selective removal of depolymerized dyed substrates from the agar replicas by solvents which neither solubilize nor precipitate the original nondegraded dyed polysaccharides present in the agar gel.
D001234:Aspergillus niger / Q000201:enzymology; D002480:Cellulase / Q000032:analysis*; D002482:Cellulose / Q000031:analogs & derivatives; D055598:Chemical Phenomena; D002621:Chemistry; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases; D006026:Glycoside Hydrolases / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology; D014990:Xylans / Q000032:analysis
D004396:Coloring Agents; D014990:Xylans; D002482:Cellulose; C002283:hydroxyethylcellulose; C044967:4-O-methyl glucuronoxylan; D006026:Glycoside Hydrolases; D002480:Cellulase; D043364:Endo-1,4-beta Xylanases