antibodies to histag proteins

Steven Cohen sbc4 at PO.CWRU.EDU
Fri Sep 13 22:18:53 EST 1996


At 12:19 PM 9/13/96 -0700, you wrote:
>Hi,
>
>I am planning to make antibodies to histag proteins without cleaving 
>the histidine tag. However, I am concerned about getting more 
>background in western blots and immunoprecipitations using antibodies 
>made to uncleaved histag proteins. Does anyone have any experience in 
>this matter ?
>
>Malini Madiraju
>
>
I am in the process of screening hybridomas generated from a GST fusion
protein. After screening 95 of them I have found 15 that are positive for my
protein, but only one this is GST positive. If you are going to go after a
polyclonal antibody, you may need to pre clear the his positive species
using some type of HIS affinity matrix.

Steve




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