Re: IP's with Decal

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From:Abizar Lakdawalla <abizarl@innogenex.com>
To:"Morken, Tim" <tim9@cdc.gov>
Reply-To:
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EDTA at that concentration (12-14%) may only go into solution after the pH is
adjusted to 8 (adjusted with sodium hydroxide). It is not very common to add
divalent ions to the EDTA reagent, as the main purpose of EDTA is to chelate and
remove divalent ions (like Ca and Mg) from the bone/cartilage.
Abizar
www.innogenex.com
1-877-IGX-INFO

"Morken, Tim" wrote:

> So how long does it take to decal with this mixture?
>
> -----Original Message-----
> From: Su, Phy-Huynh [mailto:psu@shctampa.usf.edu]
> Sent: Tuesday, August 22, 2000 5:42 PM
> To: 'Histonet'
> Subject: RE: IP's with Decal
>
> I am using EDTA buffered to decal, too.  I made it myself to have a higher
> concentration of EDTA to decal faster.  Typically I'll have 12.% to 14% EDTA
> in the solution.  Boil your water in a microwave and pour EDTA powder in.
> That will dissolve EDTA very quickly.  Cool it down, then buffer it, and add
> other stuff.  You can have your 4% paraformaldehyde, or 10% NBF in the
> solution (if you fix your tissues using those) to fix and decal at the same
> time.  I also have some MgCl2 in my decal solution.  (Only add MgCl2 after
> the solution was buffered.)  This seems to help to keep my Safranin-O
> staining for proteoglycan good.
>
> Other people decal at 37 degree, or using microwave.  This seems to speed up
> decal time.  But I have lost some antigenicity this way, so I prefer to do
> it at RT or at 4 degree with stirring.
>
> If you want to try my recipe, e-mail me.
> Good luck
> Su
>
> > -----Original Message-----
> > From: Abizar Lakdawalla [SMTP:abizarl@innogenex.com]
> > Sent: Tuesday, August 22, 2000 1:07 PM
> > To:   Jennifer Englin
> > Cc:   histonet@pathology.swmed.edu
> > Subject:      Re: IP's with Decal
> >
> > EDTA (versene) based decal solutions work best for most immuno's.
> > Downside: it's expensive and takes much more longer to decal.
> > abizar
> > www.innogenex.com
> > 1-877IGX-INFO
> >
> > Jennifer Englin wrote:
> >
> > > Has anyone found a decal that works well with immuno's?
> > > We are currently using Stephens Scientific Decal- formic
> > acid/formaldahyde/mathanol, and we are having trouble with our Kappa and
> > Lambda.
> > > Any suggestions?
> > >
> > > Jennifer,
> > > Willmar MN
> >




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