Bio-Rad RC DC™ Protein Assay - Manual (Page 4)

3 Pipet 25 µl of standards and samples into clean, dry microfuge tubes.
4 Add 125 µl RC Reagent I into each tube, vortex. Incubate the tubes for 1
minute at room temperature.
5 Add 125 µl RC Reagent II into each tube, vortex. Centrifuge the tubes at
15,000xg for 3-5 minutes.
6 Discard the supernatant by inverting the tubes on clean, absorbent tissue
paper. Allow the liquid to drain completely from the tubes.
7 Add 127 µl Reagent A´ to each microfuge tube, vortex. Incubate tubes at room
temperature for 5 minutes, or until precipitate is completely dissolved. Vortex
before proceeding to the next step.
8 Add 1 ml of DC Reagent B to each tube and vortex immediately. Incubate at
room temperature for 15 minutes.
9 After the 15 minutes incubation, absorbances can be read at 750 nm. The
absorbances will be stable for at least 1 hour.
Section 5 Storage
RC Reagent I and RC Reagent II should be stored at room temperature (20°C to
30°C) away from direct sunlight. Both reagents have a shelf life of 12 months.
(Also see DC Protein Assay Instruction Manual for storage conditions for DC
Protein Assay Reagent A, Reagent B and Reagent S.)
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