I am running WB for my protein of interest. should I use loading control together with sample during WB step or I have to do it after stripping ?
If the protein that you want to use as loading control runs at a very different apparent molecular weight than your sample protein of interest, you can simply cut the membrane after the transfer and stain each "half" with another antibody. That way you would save the trouble with stripping. Good loading controls are GAPDH (30 kDa), beta-Actin (45 kDa), calreticulin (60 kDa), or Grp94 (90 kDa). You should be able to find one that does not overlap with your sample protein.
If the protein that you want to use as loading control runs at a very different apparent molecular weight than your sample protein of interest, you can simply cut the membrane after the transfer and stain each "half" with another antibody. That way you would save the trouble with stripping. Good loading controls are GAPDH (30 kDa), beta-Actin (45 kDa), calreticulin (60 kDa), or Grp94 (90 kDa). You should be able to find one that does not overlap with your sample protein.
I agree with the Max, however, you have to be carefully to choose as loading control protein which does not changes in your samples after treatment/mutation.
In some case GADPH or actin also subjected to changes in your samples...
If you see that the usual proteins used for control are altered in your situation, you can use the rouge Ponceau staining as a loading control. This happened with me, and some researchers discuss that this would be the best control.
Can calreticulin be used as a loading control for whole cell lysates?
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