Does anyone know any good free software for measurement of protein density of western blot? I have tried "ImageJ", but there is a problem: when I select band and go in Analyse-Gels-select first lane, there is a problem with selecting second lane because it requires exactly the same size, but I cannot make the same size square.
Hi David
I use Image J and it works well by giving you a chromatogram afterwards in which you see all the protein bands represented in a peak. You select the peak you want and can copy the read out to excel. Follow the explanation in this link and it should work.
http://lukemiller.org/index.php/2010/11/analyzing-gels-and-western-blots-with-image-j/
Barbara
Dear David,
I am also using the same software,
for selecting second well just press arrow key .
Thanks
You can use Bio Rad Quantity One® 1-D software, which is user friendly and it has options for auto detection and manual detection of the Bands, in manual detection you can select the region of band to be quantified
All the best
Hi David
I use Image J and it works well by giving you a chromatogram afterwards in which you see all the protein bands represented in a peak. You select the peak you want and can copy the read out to excel. Follow the explanation in this link and it should work.
http://lukemiller.org/index.php/2010/11/analyzing-gels-and-western-blots-with-image-j/
Barbara
Hi David,
I use ROI manager in image J (under analyse) to do both weterns and PCR gels. make sure show all and edit mode are ticked. You draw your box around the band then click add. hey presto if you use your arrow keys you get another box the same size to add around your next band. Keep going as required then you can click measure and get numbers that represent the band intensity.
hope this helps.
If you are not using the arrow key, simply drag the previous box to the new band, just make sure it needs to be a straight band not up and down.
thanks for advice, I have tried with ROI (analyse-tools-ROI manager) have used some example of picture of blotting, and get
1 396 223.937 180 243
2 396 207.932 126 242
3 396 178.475 59 243
4 396 135.427 6 242
data 1 one is less intensive and 4 band was darkest.
question is how can I interpret this results? if it measures density darkest band should show highest value or?
So, How you analyze the data that you got from ROI manger, Are you dividing the ARea by the mean or what ?!!
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