A rapid method for detection of N-acetylglucosaminidase-type chitinase activity in crossed immunoelectrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels using 4-methylumbelliferyl-N-acetyl-D-glucosaminide as substrate.
A method for the detection of N-acetylglucosaminidase (GlcNAcase) activity has been developed by using 4-methyl-umbelliferyl-N-acetyl-D-glucosaminide (4-MU-GlcNAc) as substrate in crossed immunoelectrophoresis (CIE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. Visualization of the reaction with a 366 nm ultra-violet light is possible in approximately 30 min. The method is fast and sensitive in comparison with previous methods. The same band as in SDS-PAGE, showing both GlcNAcase and chitinase activity, was found in the present study; we therefore conclude that this method is also useful in a GlcNAcase-type chitinase assay.
Chen
K S
KS
Department of Marine Food Science, National Taiwan Ocean University, Keelung, Republic of China.
Lee
K K
KK
Chen
H C
HC
D000117:Acetylglucosamine / Q000031:analogs & derivatives*; D000118:Acetylglucosaminidase / Q000032:analysis*; D002688:Chitinases / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation*; D005453:Fluorescence; D005782:Gels; D006923:Hymecromone / Q000031:analogs & derivatives*; D007123:Immunoelectrophoresis, Two-Dimensional / Q000295:instrumentation*; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D013997:Time Factors
D005782:Gels; C025158:4-methylumbelliferyl 2-acetamido-2-deoxy-beta-D-glucopyranoside; D006923:Hymecromone; D002688:Chitinases; D000118:Acetylglucosaminidase; D000117:Acetylglucosamine