Table 4 Buffers and materials used for protein purification

Resin

Ni-NTA Superflow (QIAGEN)

Glutathione Sepharose 4B (Amersham Biosciences)

Amylose resin (New England Biolabs)

Resuspension/extraction

50 mM NaH₂PO₄, pH 8 300 mM NaCl 10 mM imidazole adjust pH w/NaOH

20 mM Tris-HCl, pH 8 500 mM NaCl 1 mM EDTA 1 mM DTT

20 mM Tris-HCl, pH 7.4 200 mM NaCl 1 mM EDTA

Wash

50 mM NaH₂PO₄, pH 8 300 mM NaCl 20 mM imidazole adjust pH w/NaOH

PBS, pH 7.3

20 mM Tris-HCl, pH 7.4 200 mM NaCl 1 mM EDTA

Elution

50 mM NaH₂PO₄, pH 8 300 mM NaCl 250 mM imidazole adjust pH w/NaOH

PBS 20 mM glutathione (red.)

Wash buffer 10 mM maltose