dkram at nki.nl
Mon Mar 19 09:00:24 EST 2001
We have been working with a 6His tagged protein (using Invitrogen pTRC-HisC
in top 10 bacteria) as well, the detection has been quite a problem.
Finally I succeeded with a combination of two MoAbs (Anti His6Gly from
Invitrogen 1:2500 and Anti RGS from Qiagen 1:2500) in an overnight
incubation. Since this worked well once I used these conditions in a later
experiment, not bordering about optimizing any further.
Nathan Alder wrote:
> I am currently working with protein bearing a 6X-His tag on the
> C-terminal end overexpressed from a Novagen pET system. I have been
> working on a protocol for protein detection using a hexa-His probe
> antibody with partial success. Has anyone used a 6XHis detection system
> which works with a high sensitivity? Any information/suggestions on this
> matter would be greatly appreciated.
> Nathan Alder
> UC Davis
> Life Sciences Addition
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