Thanks everyone!
Thanks Prof. David Blum. I will do batch load and check.
Yours Sincerely,
Sundaram.S
On Wed, Feb 1, 2017 at 9:06 PM, David Blum wrote:
> Hi Sundaram,
>
> The binding capacity of this column is 40 mg/mL of resin so a 5mL column
> will hold a maximum of 200 mg of protein. If y
Hi Sundaram,
The binding capacity of this column is 40 mg/mL of resin so a 5mL column
will hold a maximum of 200 mg of protein. If you run your cleared lysate
on a gel you may be able to estimate how much protein there is. Our
facility purifies a range of different proteins for investigators and
Dear Sundaram S,
during my PhD I used 4-7.5ml resin per l of culture, but I also had a large
yield of 60-100mg protein per litre. Try to use as little as possible - at
first trial check both flow-through and retentate by SDS-PAGE. (see. p. 40 and
54 of my thesis).
My constructs would also bind
This should give you everything you need to know about the column
http://www.sigmaaldrich.com/technical-documents/protocols/biology/roche/complete-his-tag-purification-column.html
I'd definitely say load all of your lysate, possibly run it through twice if
you really want to. If you're worried
