Reagents for experiments

Kang, Lin-Woo, Ph.D.Professor

Department of Biological SciencesKonkuk University

Seoul, Korea

Cell lysis buffer (25 mM Tris–HCl pH 7.5, 3 mM beta-mercaptoethanol

Digestion buffer(20 mM Tris–HCl pH 8.0, 150 mM NaCl, 3 mM -mercaptoethanol

buffer A (20 mM Tris–HCl pH 8.0, 15 mM NaCl, 3 mM -mercaptoethanol).

buffer B (20 mM Tris–HCl pH 8.0, 1 M NaCl, 3 mM -mercaptoethanol)

Crystallization buffer(20 mM Tris–HCl pH 8.0, 20 mM NaCl, 3 mM –mercaptoethanol)

YpDDL enzyme assay

50 mM Tris–HCl pH 7.810 mM MgCl210mM KClthe desired amount of D-alanine (0.1–1.0 mM to determine Km1

and 1–10 mM to determine Km2) 2 mM ATP in a total volume of 50 ul

1.5 ug YpD이

STOP10 mM EDTA (250 ul)

750 ul of Ames solution (발색반응)(0.36%(w/v) ammonium molybdate 4H2O in 1 N H2SO4 1.42% ascorbic acid)