Procedure:

1.      Dispense 2.0 mL ProteinCHEK Reagent into glass tubes.

2.      Zero or blank your instrument @595nm.

3.      Add 10 (1% of standard concentration or 10ug/ml), 20, 40, 80 and 100ul of ALerCHEK Protein Standard (1000ug/ml) respectively to each tube. A minimum of duplicates is recommended for each protein standard data point determination.

4.      Vortex immediately.

5.      Measure O.D. @595nm after 2 minutes incubation.

6.      Repeat the above using 20, 40, 60, 80 and 100 μL of standard.

*(100μL = 100% of declared value of the ProteinCHEK Standard)

7   Create a Standard Curve of O.D.@595nm (y-axis) versus protein concentration in ug/mL (x-axis).

8.      For unknowns greater than greater than 80ug/ml it is preferable to use lesser amounts of  specimen addition and multiply your answer correspondingly, e.g. 50ul addition and multiply your answer by two.

ProteinCHEK is an optimized and proprietary formulation of the original Coomassie Assay developed by Bradford, M. M. (1976) Anal. Biochem. 72, 248.

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