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Re: [ccp4bb] Tag, you're in! Flag, V5, etc |
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CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 08 February 2008Subject: Re: Tag, you're in! Flag, V5, etc From: Vangelis Christodoulou e {- dot -} christodoulou {- at -} NKI {- dot -} NL Date: 2008-02-08 An interesting approach is to add a c-term biotin tag as a bio-sensor. If the c-term of the target protein is accessible the biotin tag will get biotinylated by BirA (co-expression may be necessary depending on the expression system). You can then easily detect the target protein by WB or pull it down with streptavidin beads. I know Darren Hart is using this approach at EMBL-Grenoble. On a similar subject, OPPF is using Carboxypeptidase A to remove the c- term 6xHis tag (LysHisHisHisHisHisHis) only to leave a c-term Lysine. Have a look at: http://nar.oxfordjournals.org/cgi/content/full/35/6/e45 Hope this helps. Vangelis -- Vangelis Christodoulou Analist C Anastassis (Tassos) Perrakis lab The Netherlands Cancer Institute Dept. of Molecular Carcinogenesis - H2 Plesmanlaan 121, 1066 CX, Amsterdam The Netherlands tel: +31 (0)20 512 1951 fax: +31 (0)20 512 1954 Website: xtal.nki.nl/Tassos_group/ On Feb 7, 2008, at 22:28, Y. -F. Li wrote: > Hello, > > I'd appreciate it if anyone could provide information (experiences > or publications) on the following: > > 1. Put a tag such as FLAG, V5, etc etc, at the N- and/or C-termini, > in order for specific detection, but not interfering with protein > folding/structure; > 2. Is a linker between the tag and target protein needed? What > linkers (length, specific sequences) would you suggest? > 3. What are the choices of such tags and why would you recommend them? > > Thank you for your time and sharing in advance. > > Yongfu Li CCP4bb navigationCCP4bb <-- 2008 <-- February 2008 <-- 08 February 2008 |
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