bradford - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki
Bradford assay in microplate
Microplate analysis.
- Dilute 1mL of Bradford reagent in a total volume of 5mL with dH2O.
- Prepare a 1mg/mL BSA solution.
- Use the following concentrations for 6 wells.
Well | Concentration | Vol.BSA(uL) | Vol.H2O (uL) |
---|---|---|---|
1 | 0 | 0 | 15 |
2 | 1 | 1 | 14 |
3 | 2.5 | 2.5 | 12.5 |
4 | 5 | 5 | 10 |
5 | 10 | 10 | 5 |
6 | 15 | 15 | 0 |
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Use the rest of the wells for your samples, use 1uL of your sample and add 14 uL of dH2O (you may use duplicates/ triplicates)
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Add 200uL of diluted Bradford reagent in all the wells you used.
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Read the absorbance at 595nm in a microplate spectrophotometer
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Consult the Bradford Assay Calibration Curve:
Ex. http://www.openwetware.org/images/7/70/Calibrationcurvebradfordassay09112013zem.png