Zymography (DEG method)

Sensitivity increases by about 1,000 times, allowing for the detection of trace proteins below 0.1 ng.

- Zymography (DEG Method) When using zymography to detect new enzymes, conventional zymography has low sensitivity, making it impossible to detect trace amounts of enzymes (activity). By using SAINOME and fluorescent reagents in this system, the sensitivity increases by about 1,000 times, allowing for the detection of trace proteins below 0.1 ng. While fluorescent dyes have high sensitivity, their low molecular weight causes the increase in fluorescence due to turnover to be overwhelmed by diffusion into the gel, making detection impossible. By cutting and dividing the electrophoresis gel with SAINOME before the reaction, it is possible to prevent the diffusion of fluorescence due to turnover, allowing for accumulation and enabling the detection of trace enzyme activity. This method is called the DEG method, and many new enzymes have been discovered using it. By utilizing the DEG method, it becomes possible to comprehensively evaluate enzyme activity within samples. Comprehensive comparative evaluations between patients and healthy individuals (or between cancerous and normal tissues) can be conducted, and it is used in drug discovery target exploration. Main implementation results → Graduate School of Pharmaceutical Sciences, The University of Tokyo

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