RE: RE: Mounting Media
From: | "Plummer, Timothy B." <Plummer.Timothy@mayo.edu> |
Ian,
I put this formulation together after reading several papers on antifade
mediums and discussing mediums with tech support people from Molecular
Probes and Vector Laboratories. The formulation I described is referenced
in Transplantation, Vol 71, 132-142, No 1, 1-15-2001 as part of the
Materials & Methods. Some of the references on DABCO that I used to prepare
the formulation are: Journal of Immunological Methods, 55 (1982) 231-242
and Journal of Histochemistry and Cytochemistry, 33 No 8, 755-761, 1985.
Tim
> -----Original Message-----
> From: Dr. Ian Montgomery. [SMTP:ian.montgomery@bio.gla.ac.uk]
> Sent: Wednesday, April 25, 2001 4:22 AM
> To: histonet@pathology.swmed.edu
> Subject: Fwd: RE: Mounting Media
>
> Tim,
> Interesting mounting media that I'll give a bash. Do you have a
> reference or is it personal communication.
> Ian.
>
>
>
>
> Date: Tue, 24 Apr 2001 13:47:32 -0500
> From: "Plummer, Timothy B." <Plummer.Timothy@mayo.edu>
> Subject: RE: Mounting Media
> To: "'cmcbride@tulane.edu'" <cmcbride@tulane.edu>,
> histonet <histonet@pathology.swmed.edu>
>
> Christine,
>
> Most antibody companies (i.e., Dako, Vector) sell a medium which
> contains an
> antifade medium. Molecular Probes is also a good source. You can
> also
> prepare you own-here is my favorite formulation which also contains
> DAPI
> (Blue fluorescent nuclear counterstain), but it can be left out.
>
> My formulation for DABCO-DAPI Antifade medium is: 25 mg/mL
> 1,4-diazabicyclo[2.2.2]octane (DABCO) with 0.5mg/mL,
> 4,6-diamidino-2-phenylindole (DAPI), in 50% glycerol/phosphate
> buffered
> saline, pH 8.6. DABCO is dissolved in 100% glycerol then 2X PBS at
> pH 8.6
> is added in equal volume to the glycerol. The DAPI can be added
> from a
> higher concentration solution stock which can be stored frozen. All
> reagents can be purchased from Sigma Chemical. pH is very
> important.
>
> Tim Plummer
>
> > -----Original Message-----
> > From: cmcbride@tulane.edu [SMTP:cmcbride@tulane.edu]
> > Sent: Tuesday, April 24, 2001 10:34 AM
> > To: histonet
> > Subject: Mounting Media
> >
> > Hi Histonet,
> >
> > Does anyone know of a good mounting media to use on
> immunoflourecent
> > slides
> > with anti-fade in it?
> >
> > Christine
> >
> > Christine McBride, M.S.
> > Tulane University Medical Center
> > Center for Gene Therapy
> > SL-99
> > 1430 Tulane Ave.
> > New Orleans, LA 70112
> > cmcbride@tulane.edu
> > Phone:(504) 988-7069
> > Fax:(504) 588-5326
>
> Dr. Ian Montgomery,
> West Medical Building,
> University of Glasgow,
> Glasgow,
> G12 8QQ.
> Tel: 0141 339 8855. Extn:6602.
> Fax: 0141 330 2923
> e-mail: ian.montgomery@bio.gla.ac.uk
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