Hello, I am a beginner in enzyme purification and need help.
I want to purify amine N-methyltransferase according to the article of
Ansher & Jacoby in J. Biol. Chem. 261, pp. 3996-4001 (1986). There are
still 2 questions remaining:
1. The mentioned article gives no flow rates for column chromatography
with DEAE-Cellulose (5 x 50 cm), Hydroxylapatite (4 x 25 cm), and
Aminohxyl-agarose (1.5 x 10 cm).
2. Is it necessary to couple the protein solution to the aminohexyl-
agarose by the carbodiimide method? I intend to use EAH-Sepharose 4B
(Pharmacia). Normally this material is designed for affinity chromatography
but the paper doesn't describe any coupling procedure.
I hope that anybody can help me with advice. Many thanks in advance.
