User guide

Quantity One User Guide
15-24
Note: Relative quantity expresses the quantity of each band as a percentage of
either all the bands in a lane or the total intensity of a lane (depending on
the setting in Preferences). This can tell you that there is twice as much of
Band X than Band Y; however, it does not tell you how many micrograms
there are of Band X in the gel. Absolute quantity is calculated based on
the relative quantity, standards, and a calibration curve that you create.
First the software calculates the relative quantity of each band from the area
under the intensity profile peak. Using the Quantity Standards function, you
then identify standards of known quantity and use these to generate a
calibration curve. You can apply this curve to unknown bands in the current
image as well as other images.
To create a calibration curve, the absolute quantities of at least two bands
must be known. The greater the number of known bands and the wider the
range of their values, the more accurate the calibration curve will be.
Note: The quantities calculated by Gaussian fitting (section 14.7) cannot be used
to in conjunction with Quantity Standards. However, you can continue to
use the original trace quantities in calculating Quantity Standards after
you have Gauss-modeled your bands.
15.3.a Creating and Applying a Set of Quantity Standards
Choose Quantity Standards from the Match menu. A pop-up box will appear,
asking you to create a new curve or load a previously defined calibration
curve (if any has been saved).
Fig. 15-17. Loading a quantity calibration curve.
Selecting Create New will open the Quantity Standards dialog box.