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
  • 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)

  • Add 200uL of diluted Bradford reagent in all the wells you used.

  • Read the absorbance at 595nm in a microplate spectrophotometer

  • Consult the Bradford Assay Calibration Curve:

Ex. http://www.openwetware.org/images/7/70/Calibrationcurvebradfordassay09112013zem.png