[Histonet] Re: DAB precipitate

[Patsy Ruegg]

Oh yea, sorry I had a brain fart, I actually use the Research Genetics ready
to use DAB that is in glycerine or something so it doesn't actually freeze.
Patsy


Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: Bryan Hewlett [mailto:bhewlett <@t> cogeco.ca] 
Sent: Friday, April 21, 2006 9:26 AM
To: Patsy Ruegg; 'Pixley, Sarah (pixleysk)';
histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Re: DAB precipitate

Hi Patsy,

I have frozen aliquots of DAB for years without any problem!

Bryan

----- Original Message -----
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
To: "'Pixley, Sarah (pixleysk)'" <PIXLEYSK <@t> UCMAIL.UC.EDU>;
<histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, April 21, 2006 12:14 PM
Subject: RE: [Histonet] Re: DAB precipitate


>I thought DAB should not be frozen????
> Patsy
>
>
> Patsy Ruegg, HT(ASCP)QIHC
> IHCtech, LLC
> Fitzsimmons BioScience Park
> 12635 Montview Blvd. Suite 215
> Aurora, CO 80010
> P-720-859-4060
> F-720-859-4110
> wk email pruegg <@t> ihctech.net
> web site www.ihctech.net
>
>
> This email is confidential and intended solely for the use of the
> Person(s)
> ('the intended recipient') to whom it was addressed. Any views or 
> opinions presented are solely those of the author. It may contain 
> information that is privileged & confidential within the meaning of 
> applicable law.
> Accordingly
> any dissemination, distribution, copying, or other use of this 
> message, or any of its contents, by any person other than the intended 
> recipient may constitute a breach of civil or criminal law and is strictly
prohibited.
> If
> you are NOT the intended recipient please contact the sender and 
> dispose of this e-mail as soon as possible.
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of 
> Pixley, Sarah (pixleysk)
> Sent: Friday, April 21, 2006 7:18 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Re: DAB precipitate
>
> Dear Eva:
> I would like to hear any answers to this also. We always have a DAB 
> precipitate. To avoid it, we make the DAB and add the nickel sulfate 
> and
> H2O2 immediately before the reaction and do the reaction quickly. 
> (Using nickel in the solution increases the precipitate 
> significantly.) The longer you wait, the greater the precipitation. If 
> I am doing a lot of samples, I set half the DAB aside and change 
> solutions in the middle (add the nickel sulfate and H2O2). Otherwise, 
> I don't know any other tricks.
>
> One other phenomenon I have seen is after freezing DAB. We make up our 
> DAB solution in phosphate buffer (without H2O2 or nickel) and freeze 
> the leftover. That works very well as long as you freeze in glass 
> (i.e. new glass scintillation vials). If you freeze in some plastics, 
> I have seen a precipitate upon thawing. And the glass has to be very 
> clean--it cannot be something that was used before for DAB and then 
> bleached and cleaned. Even if you clean the glass with a good cleaner 
> like Chromerge or Contrad, there can be enough bleach leached into the 
> glass to inactivate the DAB.
>
> Good luck and I hope to hear some interesting solutions to your problem!
> Sarah Pixley
>
>
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