Protein Quantification Using ImageJ: Difference between revisions
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New page: ==Determining the concentration of protein in SDS-PAGE gel bands using ImageJ== *To determine protein concentration you will need to have a standard curve to compare your samples to ::- Fo... |
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5. Select the rectangle tool, and draw a box around the lane, '''making sure to include some of the empty gel between lanes and white space outside of the band''' (see below) | 5. Select the rectangle tool, and draw a box around the lane, '''making sure to include some of the empty gel between lanes and white space outside of the band''' (see below) | ||
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Revision as of 21:22, 29 May 2013
Determining the concentration of protein in SDS-PAGE gel bands using ImageJ
- To determine protein concentration you will need to have a standard curve to compare your samples to
- - For 5GB1, BSA works great as a protein standard, and a range of 0.025 μg/μL to 5.0 μg/μL works well as a range for the standard curve
- After running and destaining the gel, take a picture and save it as a .tif and as a .jpg (in case the tiff file can’t be opened—an issue I am experiencing at the other lab).
- -Make sure you save your images as the same type of .tif each time!
1. Download the ImageJ software: http://rsbweb.nih.gov/ij/download.html
2. Open ImageJ
3. Go to File→Open→(your image)
- Does your image look too dark or too light?
- - Image→Adjust→Brightness/contrast
- If image looks good...
4. Find the lane with the lowest concentration of BSA
5. Select the rectangle tool, and draw a box around the lane, making sure to include some of the empty gel between lanes and white space outside of the band (see below)
