Objective
- Determine concentration of ADA prepared Oct. 23 via Bradford Assay
Bradford Assay
- Solutions were prepared with 715μL 1x Bio-Rad Bradford reagant and the following concentrations of BSA:
volume BSA (uL)
|
volume H2O (uL
|
[BSA] (mg/mL)
|
[BSA] (M)
|
0 |
30 |
0 |
0.00E+00
|
5 |
25 |
0.24 |
3.61E-06
|
10 |
20 |
0.15 |
2.26E-06
|
15 |
15 |
0.13 |
2.01E-06
|
20 |
10 |
0.15 |
2.26E-06
|
25 |
5 |
0.24 |
3.61E-06
|
30 |
0 |
1 |
1.51E-05
|
- A UV/Vis calibration curve was obtained by analyzing the samples and plotting BSA concentration against absorbance at 595nm.
- The ADA purified yesterday was analyzed via UV/Vis and it's peak was compared to the linear regression of BSA's in order to determine concentration.
Data
Fraction #
|
Absorbance
|
[ADA] [mg/mL]
|
molar absorptivity [mL/mg*cm-1]
|
Molarity (µM)
|
2 |
0.93 |
0.0177 |
52.4 |
42.0
|
8 |
0.757 |
0.0115 |
65.9 |
54.4
|
9 |
0.819 |
0.0137 |
59.7 |
65.0
|
|
Frac 2 was found to be 42µM, while 8 was 54.4µM and 9 was 65µM
|