Tasks for November 5
- Enzymatic assay for lysozyme
- To carry out dialysis data analysis of solution from 11/4
Enzymatic assay for lysozyme
Preparation of 400 units/mL lysozyme
- Prepared 10mL 400 unit/mL of lysozyme
- 0.0867 g of lysozyme were added to a 10 mL volumetric flask and made up to the mark with cold phosphate buffer.
- Prepared just before addition to keep solution cold
Enzymatic assay
- Protocol followed as detailed by Sigma-Aldrich [1], with slight modifications.
Using the kinetic mode in the UV-Vis spectrometer,
- Autozero, measure a blank containing 2.0 mL of cell suspension, and add 0.1 mL of cold buffer. Record the decrease in A450 for 5 min.
- Measure a blank containing 2.0 mL of cell suspension, and add 0.1 mL of 400 units/mL lysozyme solution. Record the decrease in A450 for 5 min.
- Obtain the maximum linear rate (ΔA450/minute) for both the Test and Blank using at least a one minute interval and a minimum of 4 data points.
- Repeat protocol using 200, 300, and 350 lysozyme units/mL solutions
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