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Bradford Protein Assay
This assay allows to determine the concentration of protein in an unknown solution.
We will test the feasibility of this test with the simple-spectro using:
- BSA 2mg/mL (we have 2mL)
- commercial Quick Start™ Bradford Protein Assay from Bio-RAD.
First experiment
A. BSA 2 mg/mL diluted 2x B. Water C. Bradford protein assay diluted 5x
| Solution | A | B | C | [BSA] |
|---|---|---|---|---|
| 1 | 0 | 100 µL | 5 mL | 0.00 |
| 2 | 20 µL | 80 µL | 5 mL | 0.25 mg/mL |
| 3 | 40 µL | 60 µL | 5 mL | 0.50 mg/mL |
| 4 | 60 µL | 40 µL | 5 mL | 0.75 mg/mL |
| 5 | 80 µL | 20 µL | 5 mL | 1.00 mg/mL |
| 6 | 100 µL | 0 µL | 5 mL | 1.00 mg/mL |
Seems there is not enough BSA.
Second experiment
A. BSA 2 mg/mL B. Water C. Bradford protein assay diluted 5x
| Solution | A | B | C | [BSA] |
|---|---|---|---|---|
| 1 | 0 | 200 µL | 4 mL | 0.00 |
| 2 | 40 µL | 160 µL | 4 mL | 0.25 mg/mL |
| 3 | 80 µL | 120 µL | 4 mL | 0.50 mg/mL |
| 4 | 120 µL | 80 µL | 4 mL | 0.75 mg/mL |
| 5 | 160 µL | 40 µL | 4 mL | 1.00 mg/mL |
| 6 | 200 µL | 0 µL | 4 mL | 1.00 mg/mL |
Quick test
1 mL pure Bradfort + 0.04 mL BSA 2mg/mL:
- R: 1.36
- G: 0.64
- B: 0.57