Re: Citrate buffers

From:IHC World

Denise,

I have been using "citrate buffer" pretreatment for BrdU staining, and it
works pretty well. Untill recently I also started to use 2N HCl pretreatment
and did not see significant difference between the two pretreatments.

Concerning "sodium citrate buffer" vs. citrate buffer, I have been working
on thymine dimer antibody using both reagents and found that "sodium citrate
buffer" works noticeably better than citrate buffer although their pH values
are the same (pH 6.0). Meanwhile I also tried microwave and steamer methods,
and noticed that microwave method works slightly better than steamer. Please
note that all my tests were based on thymine dimer antibody only but thought
that BrdU is also a nuclear protein so they may have some similarities. Just
for your reference.

Richard
____________________
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----- Original Message ----- 
From: "Anna Denise R. Garcia" 
To: "IHC World" 
Sent: Friday, July 04, 2003 2:38 PM
Subject: Re: Citrate buffers


> Hi Richard,
>
> Yes, I've been trying to do double staining with bromodeoxyuridine (BrdU)
on
> GFP tissue, but the 2N HCl treatment to denature the DNA quenches the
green
> fluorescence, so someone suggested to me to try heat induced antigen
> retrieval.  Any suggestions for this type of protocol?  Some say microwave
> the tissue, some say water bath, ever done it?  Any suggestions?
>
> Thanks,
> Denise
>
>
>
> On 7/4/03 7:50 AM, "IHC World"  wrote:
>
> > For what purpose? Antigen retrieval?
> >
> > Richard
> >
> > ----- Original Message -----
> > From: "Anna Denise R. Garcia" 
> > To: "HistoNet Server" 
> > Sent: Thursday, July 03, 2003 4:43 PM
> > Subject: 
> >
> >
> >> Anyone know the difference between "Sodium Citrate Buffer" and just
plain
> >> "Citrate Buffer" or "Citric Acid Buffer"??
> >>
> >> Thanks
> >>
> >> ***************************
> >> Anna Denise R. Garcia
> >> UCLA, Department of Neurobiology
> >> Box 951763
> >> Los Angeles, CA 90095-1763
> >> (310)206-2262
> >> adgarcia@ucla.edu
> >> ***************************
> >>
> >>
> >>
> >>
> >>
> >
>
> ***************************
> Anna Denise R. Garcia
> UCLA, Department of Neurobiology
> Box 951763
> Los Angeles, CA 90095-1763
> (310)206-2262
> adgarcia@ucla.edu
> ***************************
>
>
>
>





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