Improved rapid detection of trypsin isoinhibitors using non-denaturing polyacrylamide gels with immobilised azoalbumin.

Phytochem Anal

School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8140, New Zealand.

Published: October 2011

Introduction: Proteinaceous inhibitors of animal trypsin occur naturally as isoforms in seeds and some are of interest as antinutritional or anti-pest agents.

Objective: To establish a simplified electrophorectic, in-gel method for rapid and direct detection of trypsin isoinhibitors present in crude plant extracts that are particularly suitable for many studies including rapid evaluation of cultivars.

Methodology: Azoalbumin (3%, w/v) is immobilised in 7.5% polyacrylamide gels before electrophoresis under non-denaturing conditions.

Results: This improved method eliminates the need for both time-consuming and labourious staining and destaining or renaturation steps.

Conclusion: Immobilised azoalbumin in polyacrylamide gels, run under non-denaturing electrophoresis conditions, can be used to assist rapid evaluation of trypsin isoinhibitors in numerous crude plant extracts.

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http://dx.doi.org/10.1002/pca.1290DOI Listing

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