Protein purification - (Mar/02/2007 )

Dear friends,

I have most of my protein sticking to the Ni-NTA column, how to elute it and use for the assay purpose.

Thank u

-radhu-

QUOTE (radhu @ Mar 2 2007, 05:17 AM)

Dear friends,

I have most of my protein sticking to the Ni-NTA column, how to elute it and use for the assay purpose.

Thank u

Describe your problem in detail!

-circlepoint-

go to www.qiagen.com and download the pdf for the 'qiaexpressionist'

there are troubleshooting tips in detail in this booklet. I'm guessing you have a solubility issue and will need to amend some or all of your buffers

-aimikins-

QUOTE (radhu @ Mar 2 2007, 02:17 PM)

Dear friends,

I have most of my protein sticking to the Ni-NTA column, how to elute it and use for the assay purpose.

Thank u

what do you mean by "sticking"? specific binding of His-tag to Ni, or unspecific binding beyond Ni?

-The Bearer-