[Zbrafish] HA, FLAG and GFP tags in zebrafish

Christian Broesamle via zbrafish%40net.bio.net (by Christian.Broesamle from med.uni-muenchen.de)
Thu Apr 16 04:25:27 EST 2009


Did you deyolk your embryos? If you detect the 
same non-specific bands with different secondary 
ABs, it could be that you have some very highly 
abundant, sticky proteins in your prep. Did you 
try immunostainings? You may see whether the same 
structures are stained.
Regarding tags: we detected background labelling 
on very few, but clearly stained neurons using an 
anti-myc-tag AB in non-transgenic larvae/embryos. 
Clearly, one has to be careful and do the 
approriate controls!

Christian

>Has anyone ever encountered any problems with detecting HA, FLAG or
>GFP tags in zebrafish expressing constructs containing these tags?
>
>I have used HRP conjugated anti-HA and FLAG antibodies as well as a
>non-conjugated anti-HA mouse antibody and have consistently got the
>same non-specific banding.
>
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-- 
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Christian Brösamle
Department of Biochemistry
Ludwig-Maximilians-Universität München
Schillerstrasse 44
80336 München
Germany
tel: +49 89 2180 75-451
fax:  +49 89 2180 75-415
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