I am try to blot for gluthathionylated proteins using Anti-PSSG Ab against total cell proteins. Ideally it should bind against all the proteins which are gluathionylated. Literature shows that proteins ranging from 200 to 20 Kda get glutathionylated.
But for me I am only getting few of bands of high MW proteins. Below that the bands just end abruptly without even the faintest signal. Only one time I got around 20 bands at different MWs. But after that I am not being able to get the same result. For some reason, the smaller and medium proteins are not shping glutathionylation at all. Only two bands at high MW.
Proteins are transferred properly as shown by Ponceau. Is it possible that smaller proteins are getting transferrd to the other side of the membrance?
Anybody can help?