Improved Rapid Detection of Trypsin Isoinhibitors using Non-denaturing Polyacrylamide Gels with Immobilised Azoalbumin

摘要

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