dephosphorylation - (Jun/04/2003 )
I want to know if anyone out there has a dephosphrylation method for my protein. I do not want to use Lambda - PPase since it requires a Mn++ as a cofactor to work and I do not want to use Calf Intestine or Shrimp APs since they require a heat deactivation step. I don't want my protein to be exposed to a temp. greater than 37 C and I do not want any Mn++ introduced in my assay. So if anyone out there knows something, I would really appreciate it.
You can deactivate CIAP with sodium vanadate (Na3VO4) at a final concentration of about 10 mM. Otherwise Sigma sells CIAP that is attached to agarose beads, so you can pellet it out after dephosphorylating. Good luck.