what is the equation for quantitative western blot - (Apr/29/2008 )
I'm comparing the expression of a protein in my treated animals and controls by western blot. So the way I do it is that I scan the bands ( target protein and actin), measure the color intensity in each band, then put it in this equation; target/actin in treated animals divided by target/actin in control animals. I think in this way I can have a good understanding of the net increase in my target protein. Does it make sense? Do you know any documented equation for that purpose?
What you have described is correct. I am unsure if this is documented ... Google it.
one should determine the absolute concentration of actin in each case to be sure that the treatment does not affect the actin expression
Fundamentals of densitometry:
One of the methods to quantitate the western result: http://www.lukemiller.org/journal/2007/08/...ts-without.html