User:Rebeca Rodriguez/Notebook/Chem 471/2015/09/09
|Biomaterials Design Lab||Main project page|
Previous entry Next entry
Protocol done by via Michael Bible because I'm late to class every Wednesday.
Stock solutions of HAuCl4·H2O and Lysozyme were prepared by Dr. Hartings. The procedure for their preparation can be found here.
The stock solution of HAuCl4·H2O has a concentration of 2.96 mM. The stock solution of Lysozyme has a concentration of 21.4 μM.
Sixteen 1 mL samples and ten 5 mL samples of a 45:1 Au:Lysozyme solution were prepared using the following volumes of stock solutions:
These solutions were then heated in the oven at 80°C for four hours.
A stock solution of lysozyme was created using a 10 mL volumetric flask. The aim was to make a 50 μM stock solution but the actual concentration was 62 μM.
8.88 mg of lysozyme was added to a 10 mL volumetric flask
The following 5 solutions were made using a 10 mL volumetric flask and the volumes of water and other solutions listed below.
Concentration of Lysozyme stock solution:
(0.00888 g)*[(1 mol lysozyme)/(14307 g lysozyme)] = 6.2*10-7 mols
(6.2*10-7 mols)/(0.010 L) = 62 μM
V2 = [C1V1]/C2
So, in order to make a 15 μM solution the following volume of stock solution was needed:
V62μM = (15 μM * 10 mL)/(62 μM) = 2.419 mL of stock solution
The method described above was used to determine the necessary volumes of previous serial solutions to use in each new dilution.
UV-Vis and Fluorescence spectra were collected for each of the 5 solutions that Michael created. A calibration curve for UV-Vis has been created using the absorbance at λ = 281nm. I made these beautiful graphs.
The Figure above shows the UV-Vis spectra of various concentrations of lysozyme.
The Figure above shows the calibration curve for the absorbance at λ = 281 as a function of the concentration of lysozyme in solution.
The Figure above show the calibration curve for the Fluorescence intensity integration as a function of the concentration of lysozyme in solution.
The Figure above shows the Fluorescence spectra of various concentrations of lysozyme.
Matt Hartings You need a description for how these solutions were made. Also, please list the molar extinction coefficient for lysozyme in M-1 cm-1
Michael created all of the solutions on Wednesday as I was commuting back to Georgetown.