Activity staining method of chitinase on chitin agar plate through polyacrylamide gel electrophoresis

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.

African Journal of Biotechnology Vol.4(1) 2005: 87-90