Ask help for protein purification using His-tag column!

James Bassuk bassuk at
Sun Nov 17 17:51:56 EST 2002

On 22 Oct 2002 chengzhjun at wrote:

>     my protein is about 30kDa and its pI is 5.1. I used pH=8.0 PBS binding buffer to banlance the Ni-His tag column. but when i put the protein solution into the column,the protein solution becomes turbid and the resin aggregates together and so the protein  buffer could't flow down. How can I do  with it .
>    Any information would be appreciated!
>   Carol

More info is needed:

*  What is your recombinant system?
*  How do you prepare your sample?
*  Tell me the composition of the buffer that your sample is contained.
*  Does your sample contain Calcium? (CaPO4 = insoluble)
*  Are you using IDA or NTA technology?


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