Methods Digest, Vol 81, Issue 2-'50mM tris-HEPES buffer (ph7.5)'.

Virash Gupta via methods%40net.bio.net (by virashkgupta from gmail.com)
Tue Feb 7 23:35:07 EST 2012


'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, <methods-request from oat.bio.indiana.edu>wrote:

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>   1. Tris-HEPES (Yvonne Couch)
>   2. Re: Tris-HEPES (DK)
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> Message: 1
> Date: Mon, 6 Feb 2012 15:31:15 +0000
> From: Yvonne Couch <yvonne.couch from pharm.ox.ac.uk>
> Subject: Tris-HEPES
> To: "methods from magpie.bio.indiana.edu" <methods from magpie.bio.indiana.edu>
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> 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: dk from no.email.thankstospam.net (DK)
> Subject: Re: Tris-HEPES
> To: methods from net.bio.net
> Message-ID: <QR%Xq.9509$rV2.1695 from newsfe11.iad>
>
> In article <mailman.222.1328552955.3721.methods from net.bio.net>, Yvonne
> Couch <yvonne.couch from pharm.ox.ac.uk> 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
>
>
>
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> End of Methods Digest, Vol 81, Issue 2
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-- 
Dr V K Gupta
Sr Microbiologist (Mol Biology)
IMBL, Department of Entomology
Pun. Agric. Univ., Ludhiana (Pb)-141004- India
M: 081465-55515


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