BCA Assay For Protein Concentration

updated 06/13/06 - Benjamin Costantino

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Setup:

BCA Dye - 1 part CuSO4 (Sigma C2284): 50 parts BCA (Sigma B9643)
Dilute BSA(2mg/ml) (Sigma P0834) in H2O to give concentrations of:
        0
        0.4
        0.8
        1.2
        1.6
        2.0
        2.4
        2.8

Procedure

1. 25µl of purified protein in solution + 75ul of H2O » 8µl per sample in triplicate into well of 96 well Maxisorp plate.
2. 8µl of standards in triplicate.
3. 200µl of dye in each well, mix well.
4. Incubate sealed O/N at room temp.
5. Next day, mix on plate reader before reading.
6. Read at 562nm.