6xHIS tag and immunofluorescence

roger.murphy at ludwig.edu.au roger.murphy at ludwig.edu.au
Wed Mar 1 20:58:55 EST 2000


Oddmund,

We've found that the 4-His antibody from Qiagen is very good for Western blot 
staining, better in fact than there other antibodies in our hands.

We've been expressing 6-his tagged proteins in CHO cells with mixed degree of 
success.  One protein we made in E.coli worked very well (~10% of total 
inclusion body protein) but was difficult to purify because it formed 
multimers.

Hope this helps.

Roger

In article <38BD1B3E.B34CDD5D at biokjemi.uio.no>, Oddmund 
=?iso-8859-1?Q?Nordg=E5rd?= <oddmundn at biokjemi.uio.no> wrote:
>Hello!
>
>I`m planning to make a 6xHis tagged fusion protein. Afterwards I want to use
> the
>
>tag as an epitope for immunofluorescence. Has anybody tried this? Which
> antibody
>
>do you suggest?
>
>Have anybody tried the 6xHis antibody from Clontech? Or the one from Amersham
>
>Pharmacia Biotech? Dianova? Qiagen?
>
>What is your experience with expression of His-tagged proteins in mammalian
> cell
>
>systems?
>
>Thank you very much for any help!
>
>Oddmund
>
>--
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>  Oddmund Nordgård
>  Ph.D. student
>  Institute of Biochemistry
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