RE: Alternative to Vectashield

From:"Favara, Cynthia (NIH/NIAID)"

Matt,

One of the researchers here gave me a  recipe for a mounting media called
Mowiol. I am making up and using for my aqueous IHC as it is far superior to
anything else I have used. Great resolution, dries quickly and can add a
component to reduce fading of fluorescence. I do not know how expensive it
is to make up as all the chemicals are on hand here. Should be better than
anything store bought. Might want to give this a try. Do not know if cover
slips can be removed after drying.

Mowiol

-Add 2.4g of Mowoil 4-88 [Calbiochem] to 6 g of glycerol. Stir to mix
-Add 6ml of water, leave stirring @RT for several hours
-Add 12ml of 0.2M Tris [pH8.5} and heat to 50C for 10 min with occasional
mixing
-Clarify by centrifugation @ 5,ooog for 15 minutes
-OPTIONAL: Add DABCO [1,4,-diazobicycli-[2.2.2]-octane, Aldrich] to 2.5% w/v
to reduce fading of fluorophores
-Aliquot into airtight containers and store @-20C


Stable @RT for a few weeks if in airtight tubes.

I have only used this a few times and am making up a stock for myself so I
do not know how long it can be left @RT.

Hope this is helpful!

Good Luck

Cynthia Favara



-----Original Message-----
From: Matt McElwee [mailto:MCELWEE@surgery.wisc.edu]
Sent: Thursday, May 15, 2003 9:06 AM
To: histonet@pathology.swmed.edu
Subject: Alternative to Vectashield


Hello histonetters.  
We're doing a lot of fluorescent staining on 40 micron, perfusion
fixed, vibrating microtome cut, mouse brain sections.  We're using FITC,
Rhodamine Red-X, and Cy5 conjugated secondary antibodies.  The IHC is
going pretty well, but we are going through a lot of Vectashield
mounting medium from Vector Labs.  It works well, but it's getting a bit
pricey.  Does anyone know of a cheap, alternative mounting medium for
fluorescence?  Any help would be appreciated.  Thanks.
Matt

Matt McElwee
Research Specialist
Department of Surgery
K4/617 Clinical Science Center
600 Highland Avenue
Madison, WI 53792
(608) 263-7648
mcelwee@surgery.wisc.edu



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