SDS-PAGE Prep
SDS-PAGE was prepared according to the following:
- 10 μL 0.6 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
- 10 μL 0.12 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
- 10 μL 30:1 Au/lysozyme colloid with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
- 10 μL 0.12 g/L unknown protein with 10 μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial
- Placed in heating block (set at 90 °C) for 5 minutes, and placed in the fridge overnight.
A pH 6.24 66 mM Potassium Phosphate buffer was prepared using monobasic and dibasic solids.
Dialysis Analysis
Bradford, UV-VIS, and Fluorescence analysis were run on the dialysis samples from the KI dialysis. 1 in 4 Bradford solution was prepared and the samples were prepped the same method as previously. A blank was also run. For UV-VIS and fluorescence, a 1 in 100 dilution of each was prepared and run. UV-VIS was run from 200-400 nm.
Bradford Results
Wavelength
|
Abs at 577 nm
|
Concentration (ug/mL)
|
Stock |
0.121 |
1.169902913
|
2 mM |
0.115 |
0.878640777
|
5 mK |
0.124 |
1.315533981
|
10 mM |
0.114 |
0.830097087
|
25 mM |
0.111 |
0.684466019
|
50 mM |
0.115 |
0.878640777
|
|
Fluorescence
UV-VIS
|