Any ideas of what is the problem?
I did 4 western blots, with identical results.
I load just one sample – 20 ug of protein per lane with 1:2 buffer in 8 lanes.
So the expression and quantification should be similar.
However, I keep getting reproducible results for all of the blots – last two lanes have less protein expression and different value for fractions protein/loading control. So, my quantification numbers (fractions) look like 5,6,6,5,6,6,10,11.
At first I thought I have a pipetting issue, but when I got the 4th blot with the same, different expression in the same last 2 lanes I started to think there must be something else. And it can’t be Bradford either because it is one sample!
Any ideas to this mystery?