Determining Percent Purity

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  1. Scan image and save as a tiff file (8-bit not 16-bit) with lanes oriented vertically.
  2. Open ImageJ either locally or via the webstart applet at http://rsbweb.nih.gov/ij/applets.html
  3. Using a rectangular box select entire lane.
  4. Select lane either by selecting Analyze->Gels->Select First Lane or CTRL-1
  5. Plot lane by selecting Analyze->Gels->Plot Lanes or CTRL-3
  6. Draw baseline using the line tool.
  7. If necessary use the line tool to connect peaks to the baseline
  8. Select peaks using the wand (tracing) tool
  9. Calculate purity by the area under the peak of interest divided by the area under all peaks combined.