Revised: 08/01 CHP
Summary of final step of previous procedure
50 mM Tris HCl pH 8
--Add 1 ml of PMFS (phenyl methyl sulfonyl fluride sigma P-7626 ) 100 mM which is in DMSO or isopropanol, it is a serine protease and is not soluble in water. Be careful not to get on you.
--or if a problem with serine proteases, try Roche complete protease inhibitor, one of the cat # 1-697-498
--WHY? a protease inhibitor (while native tailspike is not sensitive to proteases, if you are examining the whole cell lysis you want to see all the tailspike chains [aggregates and native]) we need to preserve the intermediates.
* Resuspend the pellet of buffer B, nothing added, to see what is in it.