Amplite® Fluorimetric Tyrosinase Assay Kit

1. Prepare and add standards and samples (50 µL)
2. Prepare and add Tyrosinase Blue working solution to the standards and samples wells (50 µL)
3. Incubate the plate at room temperature for 60 to 120 minutes
4. Monitor the fluorescence intensity at Ex/Em= 340/440 nm

Bring all the kit components at room temperature before starting the experiment.

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

Add 120 µL Assay Buffer (Component B) into Tyrosinase Standard (Component A) and mix well.

Note: Store the unused Tyrosinase stock solution at -20 °C in a single use aliquots.

Add 50 µL DMSO (Component D) into Tyrosinase Blue (Component C) and mix well. 

Note: Store the unused Tyrosinase Blue stock solution at -20 °C in single use aliquots.

Make a 1:100 dilution by adding 10 µL Tyrosinase Blue stock solution (100X) to 1 mL Assay Buffer (Component B) and mix well.

Table 1. Layout of Tyrosinase standards and test samples in a white-clear bottom 96- wells microplate. Tyrosinase standards (T1-T7= 400 to 6.25 U/mL), TS= Test Samples, BL= Blank samples

1. Prepare the standards and test samples as per recommendations in assay buffer and add 50 µL of each in a microplate.
2. Add 50 µL Tyrosinase Blue working solution to the wells of standards and samples.

3. Incubate the reaction at 37 °C for 60 to 120 minutes.

   Note: The reaction can be kept up to 6 hours.

4. Monitor the fluorescence intensity with fluorescence plate reader at Ex/Em= 340/440 nm with cutoff= 420 nm.