Protocols - Protein quantitation - BCA Method

Reagents

Standard BCA kit (Pierce #23227 or #23225)

Protocol - Standard Curve

1. The Pierce kit provides a BSA standard at a concentration of 2 mg/ml.
2. Make serial dilutions of BSA in the lysis buffer of choise.
3. Add 5-10 ul of the BSA dilutions in 1.5 ml eppendorf tubes.
4. Make enough BCA by mixing 980 ul Reagent A and 20 ul Reagent B for each sample.
5. In each tube add 1,000 ul BCA, and mix.
6. Incubate at 37°C waterbath for 30 min.
7. Transfer to plastic cuvetes.
8. Measure absorbance at 562 nm.
9. Generate standard curve based by plotting the absorbance vs the protein concentration.

Protocol - Samples

1. Make enough BCA by mixing 980 ul Reagent A and 20 ul Reagent B for each sample.
2. In a 1.5 ml eppendorf tube add 5-10 ul sample.
3. Blank tube should have lysis buffer.
4. Add 1,000 ul BCA, and mix.
5. Incubate at 37°C waterbath for 30 min.
6. Transfer to plastic cuvetes.
7. Measure absorbance at 562 nm.
8. Calculate protein concentration according to standard curve.

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