- Step:
Either load one or more equilibrium data sets with
"Load Data" or load an existing fit with "Load Fit"
- Step:
Change the project name to a sensible name (different
from equilibrium data, and no spaces!)
- Step:
Select a model by clicking on "Select Model"
- Step:
Edit noise of each scan by scrolling through all scans
and dragging the mouse over noisy data ranges. Datapoints highlighted
in red will be discarded. Reset original scan limits by clicking on
"Reset Scan Limits"
- Step:
Run "Scan Diagnostics", review excluded scans
(marked in blue) and override by double-clicking on the entry in the
listbox.
- Step:
Initialize parameters by clicking on "Initialize
Parameters". Review molecular weight settings, override
suggested values if necessary. If molecular weight is changed, re-run
"Initialize Parameters", and select "Current Molecular
Weight".
- Step:
Load or create wavelength profile, or enter extinction
coefficients manually (in OD/mol*cm), also enter buffer density and
vbar values.
- Step:
Set floatable parameters to be fitted by clicking on
"Float Parameters". Change if needed in "Model Control"
dialogue.
- Step:
Run "Check Scans for Fit" for a final check
before fitting. If successful, the "Fitting Control" button will
be activated, and can now be selected. Otherwise, review error messages
and fix.
- Step:
Select "Fitting Control". Review settings in
fitting control and select "Fit".
- Step:
After converging, save fit and attempt a fit using
the "Autoconverge" fitting mode. When saving, a default name is
provided, which should not be changed to assure all related
files can be found during project report generation. Repeat fitting until
no further improvement is observed. If program "hangs", "Abort"
fit and check amplitudes of all scans. Values should be negative or close
to zero. Also check to see if the log of the association constant is
reasonable (monomer-dimer: 8-12, trimer 18-26, tetramer: 25-35, all in
molar units).
- Step:
Close model control and select different model. Re-initialize
and repeat fitting from step 6.