Antibodies to poly-His

[Pat Thoyts]

A. Vairis wrote:
>         I'm trying to get the addresses of companies which sell any
> antibodies to a poly-His tag to immunoprecipitate an expressed protein.
> I know Qiagen do one, but I think their antibody also recognises several
> other a.a. which my construct does not contain. Any information will
> gratefully recieved,
> 
> Alan Robertson
> Dept. of Medicine
> University of Bristol
> Bristol Royal Infirmary
> Bristol BS2 8HW

True - the Quiagen antibody does recognise some extra amino acids. 
However, they are now also selling Ni-NTA alkaline phosphatase conjugate. 
This is the nickel binding molecule linked to AP. You block your filter 
and use this as for a secondary antibody and stain. It seems to work well 
on a pET - beta galactosidase controll we have - we havn't yet got around 
to using it for real. This obviously only binds to the His tag and is 
(or should be) independant of the surrounding amino acids.
The Quiagen catalogue number is #34510.

-- 
Pat Thoyts                                      thoyts at bsa.bristol.ac.uk