HELP!!!! how to purify protein require for NAD

Colin Rasmussen colin at
Mon Mar 6 11:13:50 EST 2000

ssultang wrote:

> Hi, Everyone!
> I am purifying some aldehyde dehydrogenases requiring NAD with
> 5'AMP-sepharose 4B.
> But,I could not find the activity after 5'AMP-sepharose 4B affinity
> chromatography
> In column work,
>     - starting and washing buffer : 10mM sodium phosphate buffer
>     - eluting bnuffer : containing 0.5, 3, 7,10mM NAD
>     - and at last eluting with 100mM sodium phosphate buffer.
> Where is the enzyme?
> How can I detach the guys from 5'AMP-sepharose 4B?
> Please send me your idea.
> With best regards.
> Thank you in advance for your reply......

Have you checked the flow through to make sure it even bound to the


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