User:Andrew W Long/Notebook/CHEM-671 Research/2015/09/09

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Reaction and UV-Vis Kinetic Analysis of Proteinase-K and Gold Nanoparticle Fibers & Preparation of Proteinase-K Stocks

Today proteinase-K was reacted with previously prepared gold nanoparticle fibers. Absorbance data from the reaction was taken every 2minutes for 2.5 hours in the OceanOptics Spectrophotometer.

Mass of Eppendorf tube: 1.01522 g

Mass of proteinase-K: 0.00104g (ideal value was 0.00100) Volume of water added to proteinase-K: 0.001L Molar mass of proteinase-K: 28,900g/mol Molarity of resulting solution: (0.00104g)*(1mol/28,900g)*(1/0.001L)=0.00003599M

Next we solved for the volume of the proteinase-K / water solution needed to make a 3mL 1uM final solution. M1*V1=M2*V2 => (0.00003599)*(V1)=(0.000001)*(0.003) => V1=0.00008336L = 0.08336mL

So since we know we must add 1mL of fibers, 0.08336mL of proteinase-K solution, and 0.3mL of buffer, we need 1.617mL of water to get a final solution of 3mL.

We added all components of the solution to a quartz cuvette, waited until the second recording on the OceanOptics, and added the 0.08336mL of proteinase-K (therefore the second recording is t=0 for the kinetics workup).

Chem671 09032015 AMS.png

Next, 1mg stocks of proteinase-K were prepared and stored in the freezer.

Tube Number Mass of Tube (g) Mass of Proteinase-K (g)
1 1.02157 0.00085
2 1.03162 0.00087
3 1.02798 0.00095
4 1.03275 0.00114
5 1.03057 0.00123
6 1.01392 0.00067
7 1.02938 0.00118
8 1.02917 0.00123
9 1.02993 0.00085
10 1.02223 0.00127
11 1.02055 0.00107
12 1.03129 0.00103
13 1.01359 0.00121
14 1.02997 0.00129
15 1.03117 0.00116

Matt Hartings please make that graph just lines instead of lines and points and reset the axes so that the any "changes" might be seen. Say ... y axis from 0 to 0.05