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Preprint

Pierce BCA Protein Assay Kit (23225/25227)

JB Jennifer Beck

Last updated date: Mar 31, 2024 Views: 2450

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Procedure

  1. BSA = 2 mg/ml

  2. Standard Curve: (serial dilution) 

    stdBSA (2 mg/ml)H2OValues
    510 ul0 ul2 ug/ul
    45 ul5 ul1 ug/ul
    32.5 ul7.5 ul0.5 ug/ul
    21.25 ul8.75 ul0.25 ug/ul
    10.6259.375 ul0.125 ug/ul
    Blank010 ul0 ug/ul

  3. Sample dilution:

    • 5 ul of protein in each (for duplicate reading) 

  4. Reagent dilution: (per plate)

    • 50:1 :: BCA:CuSO4. Prepare appropriate volume for 90ul per sample. 

      • BCA 

        • 1 plate = 96 wells

        • 96 wells x 95 ul = 9120ul = 9.2ml

      • CuSO4

        • (9.2 ml BCA)(1 ul CuSO4/50ul BCA) 

        • (9.2 ml)/(50 ul) = 184 ul CuSO4

  5. Incubate in dark for 30 min at 37°C. 

  6. Read in multi-reader at 570 nm.


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