Western blots are 'semi' quantitative? - (Nov/29/2011 )
Western blot is Semi quantitative because you are not measuring exact number amount of expression level of your protein.
But you can express expression in relative. To do run equal amount of protein and do western blot of your protein of interest and actin or any hose keeping protein and then measure band intensity using Imagej ( a free software you can use it measure band intensity) Then finally you can express your result that your protein of interest increase or decreased relative to the house keeping protein which you checked
Biouday on Sun Dec 11 23:44:42 2011 said:
Western blot is Semi quantitative because you are not measuring exact number amount of expression level of your protein.
But you can express expression in relative. To do run equal amount of protein and do western blot of your protein of interest and actin or any hose keeping protein and then measure band intensity using Imagej ( a free software you can use it measure band intensity) Then finally you can express your result that your protein of interest increase or decreased relative to the house keeping protein which you checked
That was what I have been doing in terms of 'quantification'. It is more of a relative 'quantification' more than anything else. For this, we have to make an assumption that the housekeeping protein is equally expressed in all cells.
if your protein of interest not interfering with housekeeping protein then the results are valid. To make sure you can check two or three housekeeping protein to get valid conclusion