Bradford Assay

From Bridges Lab Protocols
Revision as of 01:03, 4 May 2009 by Davebridges (Talk | contribs) (Reference)

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Materials

  • BioRad Protein Assay Dye Reagent Concentrate cat#500-0006
  • Disposable Plastic Cuvette

Protocol

  1. Dilute reagent 5X in water, stable for 2-3 weeks
  2. Pipet 1 mL into disposable plastic cuvette
  3. Add 1-10 uL of protein sample, cover with parafilm and mix
  4. Let sit 5-10 min to react
  5. Set spectrophotometer as follows:
    1. Go to protein assay then Bradford assay
    2. Set formula, then select more
    3. Set b=0.045 (or determine slope)
    4. Set dilution to be 1/vol (ie 0.1 for 10 uL)
    5. Blank then measure samples, absorbance must be less than 0.9
    6. Print and attach to experiment

Reference

Wikipedia: Bradford Protein Assay <pubmed>942051</pubmed>