objective
The objective for today is to begin the synthesis of BSA:L-Cysteine Nanocrystals
Preparation of stock solutions
- BSA
- L-Cysteine
- Sodium Hydroxide
- Sodium Sulfide
Protocol BSA:L-cysteine Nanocrystals
- In test tube mix 180 uL of 5mM Zinc Nitrate and 20 uL of 5mM Manganese Acetate
- Add 200 uL of 40 mM Ammonium Acetate
- Add 200 uL of Protein:ligand in various ratios from 0% to 100% protein
- Adjust pH to 8.2 using roughly 250 uL of 10mM Ammonium Hydroxide, and bring volume of water to 2.3 mL
- Incubate at room temp for 1 hour
- Add 200 uL of 5mM Sodium Sulfide drop wise while vortexing
- Incubate for five days at 37 degrees Celsius
Table of BSA:L-Cysteine ratios
mol of protein |
2.00E-09 |
1.80E-09 |
1.60E-09 |
1.40E-09 |
1.20E-09 |
1.00E-09 |
8.00E-10 |
6.00E-10 |
4.00E-10 |
2.00E-10 |
0
|
percentage of protein |
100 |
90 |
80 |
70 |
60 |
50 |
40 |
30 |
20 |
10 |
0
|
percentage of ligand |
0 |
10 |
20 |
30 |
40 |
50 |
60 |
70 |
80 |
90 |
100
|
mol of ligand |
0 |
2.00E-10 |
4.00E-10 |
6.00E-10 |
8.00E-10 |
1.00E-09 |
1.20E-09 |
1.40E-09 |
1.60E-09 |
1.80E-09 |
2.00E-09
|
|