Bradford Assay
Bradford assays are used to determine the concentration of a protein by comparing the optical density of the protein to a standard curve.
Contents
Section titled “Contents”1. Standard curve
Section titled “1. Standard curve”To make the standard curve follow the table below:
| Standard | Bovine Serume Albumin (uL) | Comassie Reagent (mL) | ddH20 (uL) | Concentration ug/mL |
|---|---|---|---|---|
| A | 30.00 | 1.5 | 0.0000 | 2000 |
| B | 22.50 | 1.5 | 7.5000 | 1500 |
| C | 15.00 | 1.5 | 15.000 | 1000 |
| D | 11.25 | 1.5 | 18.750 | 750 |
| E | 7.500 | 1.5 | 22.500 | 500 |
| F | 3.750 | 1.5 | 26.250 | 250 |
| G | 1.875 | 1.5 | 28.125 | 125 |
| H | 0.375 | 1.5 | 29.625 | 25 |
| I | 0.000 | 1.5 | 30.000 | 0 |
Use the Nanodrop to make the standard curve. It should be linear.
2. General Protocol
Section titled “2. General Protocol”- Blank the nanodrop by depositing 1 mL of H2O into a cuvette.
- Place the cuvette in the machine (clear windows following the arrow) and click blank.
- Now measure each standard (A-G). Note: Re-use the same cuvette for all standards, starting with most dilute first.
- Once done, measure your sample.
- Throw away cuvettes.