[박상규] buffers for p43 purification

buffers for p43 purification

Column Binding buffer (500ml)

20 mM KH2PO4, 500mM NaCl, pH 7.8, 2mM b-mercaptoethannol

Column Wash buffer 1(500ml)

20 mM KH2PO4, 500mM NaCl, 10% glycerol, pH 6.0, 2mM b-mercaptoethannol

Column Washbuffer 2(500ml)

20 mM KH2PO4, 500mM NaCl, 10% glycerol, pH 5.2, 2mM b-mercaptoethannol

column wash buffer 3 (500ml)

20 mM KH2PO4, 500mM NaCl, 10% glycerol, pH 6.0, 2mM b-mercaptoethannol + 100mM Imidazole

3M imidazole in D.W

for Elution

Dilute 3M imidazole to 300mM with column wash buffer 1

1X PBS containing 20% glycerol for dialysis

è DW for buffer should be 3rd. Solution for purification should be sterilized by filtration of 0.2um membrane and adjust pH with HCl not dibasic phosphate. Mercaptoethanol should be added just before use