- We took the absorbtivity of lysozyme with UV VIS and Fluorescence More detailed instructions can be found here
- Lysozyme samples for spectral analysis
- Lysozyme has a molecular weight of 14307 g/mol
- We prepared a 51.2uM lysozyme stock soluton by weighing out 7.32 mg of lysozyme and transfered it to a 10 mL volumetric flask where we pipetted to the mark with deionized water. We transfered our stock solution to a falcon tube.
- We made 5 lysozyme samples for analyzing with UV Vis and fluorescence spectroscopies between a working range on 0 and 15 uM.
- (volume of stock or previous sample)(Conc of used sample
0/(10 mL deionized water)=new sample conc.
- (2.93 mL)(51.2 uM)/10 mL= 15 uM
- (5 mL)(15 uM)/10 mL= 7.5uM
- (7 mL)(7.5 uM)/10 mL=5.25 uM
- (5 mL)(5.25 uM)/10 mL= 2.625 uM
- (3 mL)(2.625uM)/10 mL= 0.7875 uM
- UV Vis data
- We measured a water blank
- We ran our 5 samples within the working range, using the UV Vis and Flurescence.
- Correct the spectra for solvent and baseline
- Flurescence data
- Measure a water blank
- Measure the spectrum of each of your samples (except for your 50 uM sample)
- Using Excel, calculate the area underneath the curve (i.e. integrate) for each of your spectra.
- Prepping protease samples
- Record the mass of 1 eppendorf tube (This is important. We will use this data for something later on) and then TARE the balance
- Add roughly 1 mg of your protease to the tube and record the mass
- Label the tube with the name of the protease and what the concentration will be after you add 1 mL of water.
- Place in your freezer box
- Processing the fiber samples
- Centrifuge the fiber samples
- Carefully draw off the liquid supernatant (without disturbing the fibers). Its OK if you leave some water on the fiber.
- Leave the tubes open so that the water can evaporate.
- We will analyze these samples on a later date.
Matt Hartings Change the A vs Conc graph so that your data shows up as points, not as a curve. What is the line fit that you made? What is your calculated molar absorptivity in M-1 cm-1