赛默飞世尔Thermo Fisher X10 HYBOND-

Phosphoamino acid analysis:Mark Kamps's method

Recipes for the buffers are at the end of this protocol. 1.Label your protein with 32Pi.Then subject the protein to SDS polyacrylamide gel electrophoresis and transfer your gel-fractionated protein to Immobilon-P. Neither nitrocellulose nor nylon

Colony Hybridization

bacteria Saran wrap Denaturing solution (0.5 M NaOH + 1 M NaCl) Neutralizing solution (1 M Tris pH 7.4, 1 M NaCl) 2X SSC, 0.1% SDS 83 mm circular nylon membranes (Hybond N) Stratalinker UV box Radiolabled probe

Two dimensional peptide mapping

so that you have at least 500 cpm. Transfer the protein to nitrocellulose, Immobilon or Nylon according to Kamps's Western Protocol . Tryptic digestion can be done with either NC, Nylon, or Immobilon. Nitrocellulose seems to give more reproducible yields of peptides.