Quantification of proteins in Western Bands
Posted 06 April 2011 - 07:45 AM
I have a protein in my X-ray film that degrades or disappears over time. I am trying to quantify the intesity of the bands over time for example, by what percentage the protein band decreases before finally disappearing. I am using ImageJ (the one found in NIH), but no luck in actually getting a good graph with the numbers I get. Any help will be appreciated.
Posted 13 April 2011 - 07:10 AM
I also use ImageJ. It is not as easy as I thought it might be. Do you run a standard together with your sample? That is the best way. I have found that you should try an dilute the sample such that it is not to dark. It appears as though the software reaches a saturation limit quite soon. I normally run 4 dilutions of standard followed by 5 dilutions of the protein. I then draw a standard curve and plug in the values for the sample(s).
Posted 25 February 2012 - 09:17 PM
I've had good results with ImageJ for western blots, HUVEC migration, and cell counting. I follow this tutorial for western blot densitometry: http://howtowesternb...quantification/
Edited by PhilB2442, 25 February 2012 - 09:18 PM.