User:Benjamin Friedel/Notebook/CHEM 471/2015/09/15

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AuNP Lysozyme Fiber Preparation

Objective

The objective is to use previously prepared stock solutions to generate AuNP fibers (AU: Lysozyme ratio = 45) for further experiments with lysozyme.

Procedure

Using the same procedures as on 09/02/2015 two sets of AuNP fiber reaction mixes were set up, one with 1 mL samples and the second with 5mL samples. Below are the contents and volumes of each reaction mix with 16x1mL samples and 10x5mL samples. All reaction mixes were prepared at room temperature with stock solutions prepared by Dr. Hartings previously the same day (09/15/2015). Samples were covered in aluminum foil then heated in oven by Dr. Hartings for 4 hours at 80˚C. The reaction cycle also included 30 minutes to heat from room temperature to 80˚C and 30 minutes to return to room temperature before and after the 4 hour reaction. This is the second attempt at AuNP synthesis in 5mL reaction mix volumes, as the first attempt on 09/02/2015 failed (for both the 1mL and 5mL samples).


Note: The final concentration of gold should be 0.25 mM and the final concentration of lysozyme should be 5.556 μM. This gives a Au:lysozyme ratio of 45, which should yield fibers.

  1. 1 mL samples
    1. Volume of gold stock: 84.4 μL
    2. Volume of protein stock: 276 μL
    3. Volume of water: 640 μL
  2. 5 mL samples
    1. Volume of gold stock: 422 μL
    2. Volume of protein stock: 1,380 μL
    3. Volume of water: 3,198 μL

Data

Group UV-Vis Data

  1. Below is a table that includes the group UV-Vis Data at 280nm for various lysozyme concentrations
Absorbance at 280nm ' '
Lysozyme Concentration (uM) A(280)
Group 1 2 0.052
5 0.16
8 0.239
10 0.292
15 0.444
Group 3 0.7875 0.026
2.625 0.091
5.25 0.186
7.5 0.252
15 0.502
Group 4 0.965 0.032
1.93 0.069
3.86 0.123
7.225 0.261
15.445 0.517
Group 5 1.56 0.052
3.13 0.108
6.25 0.221
12.5 0.441
15 0.54

Figure 1, below, shows the calibration curve for the group UV-Vis Data for Lysozyme at 280nm. From this equation, we can see that the molar absorptivity is 33.2Lmol-1cm-1.

Image:09.15.2015 Group Data JNB Lysozyme UV-VIS.png

Tube Data

  1. 1.5mL eppendorf tubes were massed by all five groups so achieve a large sample size to calculate the average mass of an eppendorf tube (same brand and volume). This will be used to calculate the mass of synthesized AuNP fibers in the future.
AMS 9/9 AMS 9/15 LMN 9/9 JBN 9/9 MB 9/9 RAM 9/15 AMS 9/16
1.02157 1.03107 1.03104 1.015 1.026 1.03005 1.03028
1.03162 1.03192 1.02859 1.01348 1.026 1.0255 1.02355
1.02798 1.02697 1.0199 1.0179 1.026 1.02676 1.02919
1.03275 1.01923 1.02753 1.02954 1.026 1.02869 1.02559
1.03057 1.02998 1.01646 1.0228 1.026 1.01656
1.01392 1.03059 1.03078 1.02048 1.026 1.01995
1.02938 1.02025 1.02419 1.01945 1.026 1.01707
1.02917 1.02932 1.01308 1.02723 1.026 1.02417
1.02993 1.03331 1.02896 1.02951 1.026 1.02518
1.02223 1.0287 1.0288 1.01517 1.026 1.01762
1.02055 1.02554 1.02602 1.02954 1.026 1.0349
1.03129 1.01372 1.02456 1.03211 1.026 1.03024
1.01359 1.01853 1.01719 1.0192 1.026 1.02016
1.02997 1.02051 1.02863 1.02781 1.026 1.0205
1.03117 1.03015 1.0285 1.026 1.02812
1.01972
Average Mass (g) Standard Deviation
1.025135426 0.005373647

Results

  1. 5mL reactions all yielded fibers (10/10) and 1mL reactions yielded no fibers, but color change from gold to purple indicated AuNP formation.

Matt Hartings What are the class's UV-Vis and tube data?