Yes, sucrase is the same as invertase.
Due to the high sugar content in honey, a 10 kDa cut-off centrifugal filter is needed in this experiment to remove background sugars (e.g. VWR Cat No: 82031-348).
1.) Accurately weigh out 10 mg of honey and dissolve it in 500 µL of H2O (50 fold dilution).
2.) Add 500 µL of the dissolved honey into the centrifugal filter. Centrifuge at 12,000 × g at 4°C until most of the liquid (there should be less than 50 µL left) has passed through the membrane (~20 min). Dispose of the eluent.
3.) To wash the invertase, add 500 µL of H2O to the tube and centrifuge at 12,000 × g at 4°C. Dispose eluent. Repeat wash one more time. Allow most of the liquid (less than 50 µL should remain) to pass through the filter.
4.) Dilute the concentrated invertase in the filter membrane by adding 500 µL of 1X reaction buffer. The sample is ready to be assayed. Refer to the EIVT product datasheet for instructions.
Note: Levels of invertase may vary in honey samples, therefore if the signal is to high, serially dilute the invertase sample to determine the appropriate dilution to work with and multiply the calculated activity by the dilution factor. If the signal is too low, adjust the dilution in step 4.
Adjusted Calculation: Invertase Activity: ((Rsample – Rcontrol)/(slope × t)) × (5×10-4 L/10 mg) (U/mg) Where 5 ×10-4 L comes from the 500 µL of reaction buffer added to dilute the concentrated invertase in the membrane and 10 mg was the initial mass of honey.
Chua, L. S., et al (2014). Effect of thermal treatment on the biochemical composition of tropical honey samples. International Food Research Journal 21(2): 773-778. Assay: Invertase in honey.
Chua, L. S., et al.(2014). Effect of thermal treatment on the biochemical composition of tropical honey samples. International Food Research Journal 21(2): 773-778. Assay: Invertase/Sucrase in Honey.
Ciesielska, K et al (2014). Exoproteome analysis of Starmerella bombicola results in the discovery of an esterase required for lactonization of sophorolipids. Journal of Proteomics(98):159-174. Assay: Invertase in S. bombicola protein extract.
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