6. Analyzing Western Blots Using ImageJ Software

Post date: May 26, 2018 4:59:07 PM

1. Open image through the software. File àOpen (as TIF/JPEG)

2. ImageàType à8-bits

3. ProcessàSubtract background (use default 50.0 pixels)

4. AnalyzeàSet measurements (select Area, Integrated Density, and Standard Deviation)

5. AnalyzeàSet scale (select pixels, which is default) àOK

6. EditàInvert image

7. Choose Rectangle from toolbar

8. Select area you want to measure (keep it tight!)

9. Hit M on the key board after selecting the area to see the measurements

10.Save the table (will open in Excel)

11.Normalize the intensity:

a. Divide IntDen for protein of interest by IntDen for beta-actin or house-keeping gene