Protein assay for samples with DTT

1. Label sufficient microfuge tubes for 6 standards and samples.

2. Add 0, 5, 10, 15, 20, and 25 ul of BSA standard (concentration of stock is 2mg/ml). Add appropriate amount of samples to tubes. Bring all standards and samples to 500 ul with water.

3.Add 50ul of 0.15% Deoxicholate to each tube. Vortex and sit at RT for 5 min.

4. Add 35 ul of 100% TCA to each tube. Vortex and sit at RT for 5 min.

5.Centrifuge each tube in microfuge, full speed, for 5 min. Aspirate off supernatant.

6. Add 50 ul of 1% SDS to each tube. Vortex

7. Add 1 ml of BCA mix to each tube. Vortex and incubate at 370 C for 30 min. Vortex frequently during incubation.

8. Allow tubes to cool to room temperature. Read absorbance at 562 nm.