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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