Hi, I have used TEV protease in the past, it is good and easy to produce at large amounts. However I prefer for some cases thrombin, it has higher activity and efficiency especially in hard conditions for TEV protease. Cleavage site: Leu-Val-Pro-Arg-Gly-Ser
You can buy it cross linked to agarose, it means you incubate it and recollect it for future use (too many times!!), just like Ni-NTA. Bit expensive but reusable at least ten times. Thrombin cleavage kit, Cat: RECOMT-1KT) Regards toufic el arnaout On Tue, Oct 9, 2012 at 6:04 PM, <[email protected]> wrote: > Send Methods mailing list submissions to > [email protected] > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/methods > or, via email, send a message with subject or body 'help' to > [email protected] > > You can reach the person managing the list at > [email protected] > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Methods digest..." > > > Today's Topics: > > 1. Re: Recombinant protein with specific cleavage in fungi > (Cathal Garvey) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sat, 06 Oct 2012 22:34:29 +0100 > From: Cathal Garvey <[email protected]> > Subject: Re: Recombinant protein with specific cleavage in fungi > To: [email protected] > Message-ID: <[email protected]> > Content-Type: text/plain; charset=ISO-8859-1 > > Hi DK, > Is there any reason not to recommend enterokinase? Recognition site is > DDDDK (right up your alley, no? ;)). I'm working on a project of my own > and had been planning to use this enzyme instead. > > On 04/10/12 05:27, DK wrote: > > In article <[email protected]>, [email protected](DK) > > wrote: > > &species= > >> Paper describing the construct and outlining purification: > >> 17543538 > > > > Sorry! That number is Pubmed ID. > > We skip all the fancy equipment in the paper and just do > > gravity column with SP-Sepharose and step elution followed > > by gravity Ni-NTA and also step elution. > > > > > > > > > > > > > > _______________________________________________ > > Methods mailing list > > [email protected] > > http://www.bio.net/biomail/listinfo/methods > > > > -- > www.indiebiotech.com > twitter.com/onetruecathal > joindiaspora.com/u/cathalgarvey > PGP Public Key: http://bit.ly/CathalGKey > > > > ------------------------------ > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > > End of Methods Digest, Vol 89, Issue 4 > ************************************** > > _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
