Anti His-tag antibodies in indirect immunofluorescence?

olivier.leger from merckgroup.com via methods%40net.bio.net (by olivier.leger from merckgroup.com)
Tue Dec 6 11:45:09 EST 2011


You should try the following reagent which works well in our hands at 1 
ug/ml
 polyHistidine antibody (monoclonal Mouse IgG1 clone # AD1.1.10)       R&D 
systems cat# MAB050 

Olivier 


From:   dk from no.email.thankstospam.net (DK)
To:     methods from magpie.bio.indiana.edu
Date:   06-12-11 17:08
Subject:        Anti His-tag antibodies in indirect immunofluorescence?
Sent by:        methods-bounces from oat.bio.indiana.edu



I need to find out localization pattern for a 8His-tagged protein. 
(This came up unexpectedly; had I planned it, there would be 
a good tag, 3HA/Flag/Myc).  My understanding is that all existing 
anti-polyHis monoclonals are rather low affinity. Yet, now there 
seems to be a bewildering variety of various anti His-tag Ab 
commercially available. Anyone has hands on experience with 
using them in indirect immunofluorescence? 

Any comments, sources, typical dilutions that don't stain 
everything , etc? They are pricey so I can't try everything and 
find out what works best. 

Thanks!

- Dima
 
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