Ask help for protein purification using His-tag column!

[James Bassuk]

On 22 Oct 2002 chengzhjun at hotmail.com 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
>
>
> http://biowww.net/mynews/tree.php?group_name=bionet_molbio_proteins&begin=0
>

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?

Jim