Hello Scientists, RFU is negative in trypsin and fluorogenic substrate Z-RR-AMC fluorogenic assay What is the reason?

02 April 2024 1 5K Report

I am performing fluorogenic assay with trypsin and fluorogenic assay Z-RR-AMC, my readings are decreasing with increasing time. What could be the reason for this strange results. Please guide.

Thanks in advance

Adam B Shapiro

Excessive illumination of the the AMC fluorophore could result in photobleaching, which results in a loss of fluorescence over time. If you are using a continuous source of illumination, reduce the beam intensity by reducing the slit width on the excitation side. If you are using a flash lamp-type system, reduce the number of flashes per reading, the frequency of readings, and/or limit the duration of the measurement.

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