Bio-Rad Protein Assay - Manual (Page 9)

7
3. Pipet 10 µl of each standard and sample solution into separate
microtiter plate wells.
4. Add 200 µl of diluted dye reagent to each well. Mix the sample and
reagent thoroughly using a microplate mixer. Alternatively, use a
multi-channel pipet to dispense the reagent. Depress the plunger
repeatedly to mix the sample and reagent in the well. Replace with
clean tips and add reagent to the next set of wells.
5. Incubate at room temperature for at least 5 minutes. Absorbance will
increase over time; samples should incubate at room temperature for
no more than 1 hour.
6. Measure absorbance at 595 nm.
Microassay Procedure for Microtiter Plates
1. Prepare three to five dilutions of a protein standard, which is repre-
sentative of the protein solution to be tested. The linear range of the
assay is 8.0 µg/ml to approximately 80 µg/ml. Protein solutions are
normally assayed in duplicate or triplicate.
2. Pipet 160 µl of each standard and sample solution into separate
microtiter plate wells.
3. Add 40 µl of dye reagent concentrate to each well. Mix the sample
and reagent thoroughly using a microplate mixer. Alternatively, use a
multi-channel pipet to dispense the reagent. Depress the plunger
repeatedly to mix the sample and reagent in the well. Replace with
clean tips and add reagent to the next set of wells.
4. Incubate at room temperature for at least 5 minutes. Absorbance will
increase over time; samples should incubate at room temperature for
no more than 1 hour.
5. Measure absorbance at 595 nm.
LIT33C 8/31/98 02:25 PM Page 7