biochrom ultrospec 7000 User Manual page 47

1. Mix 50 parts of Reagent A (a solution containing bicinchoninic acid, sodium
carbonate, sodium tartrate and sodium bicarbonate in 0.1 N NaOH, pH 11.25) with
1 part of Reagent B (4% (w/v) CuSO
standards and samples. 2 ml of working reagent is required for each sample.
2. Mix until the solution is a uniform light green color. The solution is stable for 1 day.
Standard Preparation
1. Prepare a series of protein standards ranging in concentration from 0.2 to
1.0 mg/ml such that the final volume for the assay is 0.1 ml. The Ultrospec
7000/8000/9000 can measure up to 9 standards and up to 3 replicates.
2. Add 2.0 ml of the BCA working reagent to each standard, vortex gently and
incubate using one of the following parameters: 60°C for 15 minutes, 37°C for
30 minutes or room temperature from 2 hours to overnight.
3. If required, allow the tubes to cool to room temperature.
Sample Preparation
1. Prepare the unknown samples as described above ensuring that the final volume
is 0.1 ml.
2. Add 2.0 ml of the BCA working reagent to each sample, vortex gently and
incubate using one of the following parameters: 60°C for 15 minutes, 37°C for
30 minutes or room temperature from 2 hours to overnight.
3. If required, allow the tubes to cool to room temperature.
Creating A Standard Curve
For BCA measurements Wavelength is set to 562.0 nm. Bandwidth (Ultrospec 9000
only), Integration Time and Lamp Mode and Sample can be set as you wish.
You may advance to the next screen at any time by pressing the forward arrow
and return to the previous screen by pressing the back arrow.
Set Calibration to Standards, Curve Fit to Zero Regression and enter Units of
mg/ml. The number of standards and replicates can be set as you wish (for
optimum accuracy it is recommended that the number of standards is ≥4 and
replicates is >1).
Note: With Calibration set to Standards the user is required to prepare and measure
standards with Calibration set to Manual the user inputs both the standard
concentrations and absorbances.
Enter the concentration of prepared standards using the numeric keypad.
5061-046 Rev1
29004782UM AA 10/2011
.5H O), preparing sufficient reagent for all the
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