ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2005
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2005 International Scholars Journals
Full Length Research Paper
Activity staining method of chitinase on chitin agar plate through polyacrylamide gel electrophoresis
Vipul Gohel, Pranav Vyas and H.S. Chhatpar*
Department of Microbiology and Biotechnology Centre, Faculty of Science, M.S. University of Baroda, Vadodara - 390 002, Gujarat, India.
Accepted 15 November, 2004
Abstract
A method for detection of chitinase activity on chitin agar plate after polyacrylamide gel electrophoresis is described. Different staining dyes such as calcofluor white M2R, fluorescein isothiocyanate, rhodamine B, ruthenium red and congo red were separately incorporated in chitin agar plates. After running polyacrylamide gel electrophoresis, the gel was transferred onto chitin agar plate containing different dyes for the activity staining. Thin layer of acetate buffer (0.2 M, pH 5) was pored on the gel, which helps faster diffusion of the enzyme from gel onto the plate. After incubation of about 7 h, bands of chitinase were visible by daylight or UV light. The method is very sensitive since it can detect even 0.5 units of chitinase. Thus, this method is sensitive, rapid, user-friendly, reliable and cost effective.
Key words: Activity staining, chitinase, dyes, sensitivity, stability.