'50mM tris-HEPES buffer (ph7.5)'. Suspend 6.055 g tris base in one litre MQ water and adjust pH to 7.5 by adding solid HEPES by continuousely mixing by keeping the container on a magnetic stirrir. This will take quite a time to become stabilized. HEPES is an acidic component and will substitute HCL used for making Tris CL (to neutralize Tris base)
On Tue, Feb 7, 2012 at 10:37 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. Tris-HEPES (Yvonne Couch) > 2. Re: Tris-HEPES (DK) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Mon, 6 Feb 2012 15:31:15 +0000 > From: Yvonne Couch <[email protected]> > Subject: Tris-HEPES > To: "[email protected]" <[email protected]> > Message-ID: > <fc9d052c4a50c04e8d6c54d2f870a5819380041...@exmbx01.ad.oak.ox.ac.uk > > > Content-Type: text/plain; charset="us-ascii" > > Hi all, > I am trying to replicate the experiments of an ex-colleagues whose notes > require '50mM tris-HEPES buffer (ph7.5)'. I am not sure whether the 50mM > refers to the tris or the HEPES and was wondering whether anyone had any > experience with this? I am using it as an extraction buffer for fresh > tissue (the other components of which are sucrose and EDTA) and need to > maintain the integrity of the enzymes within the tissue, rather than the > cells. If anyone has any suggestions for a replacement for this buffer or > any more details on how to make a tris-HEPES buffer, I would be most > grateful. > Regards > Yvonne > > > > ------------------------------ > > Message: 2 > Date: Tue, 07 Feb 2012 02:18:25 GMT > From: [email protected] (DK) > Subject: Re: Tris-HEPES > To: [email protected] > Message-ID: <QR%[email protected]> > > In article <[email protected]>, Yvonne > Couch <[email protected]> wrote: > >Hi all, > >I am trying to replicate the experiments of an ex-colleagues whose notes > > require '50mM tris-HEPES buffer (ph7.5)'. I am not sure whether the 50mM > > refers to the tris or the HEPES and was wondering whether anyone had any > > experience with this? > > There is plenty of ambiguity there but I'd think that it means that > the buffer is made with 25 mM of Tris and 25 mM of HEPES. > At least that's how it works in our lab: a "1.0 M MES-Acetate" > is 0.5 M of each component. > > > has any suggestions for a replacement for this buffer > > It's going to be a very good buffer at 7.5. Lots of things could > work as replacement though. > > >or any more details on > > how to make a tris-HEPES buffer, I would be most grateful. > > Mix HEPES (free acid form) and Tris (free base form) and > adjust pH to 7.5 with either NaOH or HCl (I am not sure what > pH is going to be when mixing equal concentrations of > HEPES and Tris). > > DK > > > > ------------------------------ > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > > End of Methods Digest, Vol 81, Issue 2 > ************************************** > -- Dr V K Gupta Sr Microbiologist (Mol Biology) IMBL, Department of Entomology Pun. Agric. Univ., Ludhiana (Pb)-141004- India M: 081465-55515 _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
