Hi, incubation of HMGB1 in 2% formic acid at 100 degrees for 20 minutes cleaves HMGB1 between A- and B-boxes.

Thanks, I appreciate your replay

I woundering Did you try it before and what the predicted Molecular wieght for each box after running western blotting

I have tried it before: sizes are 15 kDa (B-box + acidic tail) and 10 kDa (A-box).

Thank you very muck for ur kind help

Hi Dr. Ari

as you suggest using formic acid,i still have the problem of having the acidic tail with the B box.

another inquiry, after cleavage of full length protein how to purify both box separately

waiting ur kind replay

Hi Mohamed,

for purification you can use heparin-Sepharose or ion exchange chromathographies.

Ari

Thanks for your kindness