Protein Quatitation by Bradford Method - Code E531, E532, E530

Directions for use

 

Components: E531 1,5ml BSA (0,5mg/ml)
a E532 200ml 0.15N NaCl
a E530 1L Bradford Reagent
Storage Cold a

 

Instructions:

  • Into 4 separate microcentrifuge tubes, aliquot 5, 10, 15 and 20 l of  0.5 mg/ml BSA solution.  Bring the volume of each to 100 l with 0.15N NaCl.

  • Into 1 tube, aliquot 100 l  0.15N NaCl.  This will serve as a blank;
  • Add to each tube, 1 ml Bradford Reagent and vortex.  Allow to stand at room temperature for 2 minutes.
  • Determine A595 nm using a 1 ml microcuvette.  Generate a standard curve by plotting absorbance at 595nm versus protein concentration.
  • For the unknown sample, repeat step 1-4 using the unknown in place  of the BSA.  Plot the A595 nm and use the standard curve as a  reference to determine the concentration of the unknown sample.

 

If after the initial assay, the unknown protein concentration is too high, dilute the protein or assay a smaller aliquot of the unknown

*Bradford, M.M. 1976.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal. Biochem. 72:248-254.

 

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