Removing DAB IHC staining

[Paul Rigby-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi All,
One of our users has had a mishap with some very precious tumour sections which were accidentally stained with the wrong primary antibody followed by DAB. They want to know if it is possible to destain the sections (remove the DAB, which is, apparently, quite dark). They have experience removing immunofluorescence antibody staining but not the DAB product.
All suggests will be very gratefully accepted.
Many thanks
Paul

Assoc. Prof. Paul Rigby
Optical Microscopy Specialist
Centre for Microscopy, Characterisation & Analysis (M510)
The University of Western Australia
35 Stirling Highway
Crawley  WA  6007
Australia

[Michael Schell-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

The only thing I know that works for this is fish.  Some breeds of tiny fish find the DAB precipitate to be quite the delicacy.  Fill your Coplin jar with the purest triple-distilled water, then add fish.  You need the type used in toe salons.  It will take a while, so be sure to change the fish frequently.  The fish must be very hungry for this to work.

> On Apr 1, 2015, at 1:08 AM, Paul Rigby <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
> One of our users has had a mishap with some very precious tumour sections which were accidentally stained with the wrong primary antibody followed by DAB. They want to know if it is possible to destain the sections (remove the DAB, which is, apparently, quite dark). They have experience removing immunofluorescence antibody staining but not the DAB product.
> All suggests will be very gratefully accepted.
> Many thanks
> Paul
>
> Assoc. Prof. Paul Rigby
> Optical Microscopy Specialist
> Centre for Microscopy, Characterisation & Analysis (M510)
> The University of Western Australia
> 35 Stirling Highway
> Crawley  WA  6007
> Australia

[Smith, Benjamin E.]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

I found this post where apparently someone managed to accidentally destain their DAB stained tissues.  Sounds like a bit of triton-X at 4oC over a weekend does the trick: http://www.ihcworld.com/smf/index.php?topic=666.0

-Ben Smith

Benjamin E. Smith, Ph.D.
Samuel Roberts Noble Microscopy Laboratory
Research Scientist, Confocal Facility Manager
University of Oklahoma
Norman, OK 73019
E-mail: [hidden email]
Voice   405-325-4391
FAX  [phone]
http://www.microscopy.ou.edu/
________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Michael Schell [[hidden email]]
Sent: Wednesday, April 01, 2015 8:47 AM
To: [hidden email]
Subject: Re: Removing DAB IHC staining

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

The only thing I know that works for this is fish.  Some breeds of tiny fish find the DAB precipitate to be quite the delicacy.  Fill your Coplin jar with the purest triple-distilled water, then add fish.  You need the type used in toe salons.  It will take a while, so be sure to change the fish frequently.  The fish must be very hungry for this to work.

> On Apr 1, 2015, at 1:08 AM, Paul Rigby <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
> One of our users has had a mishap with some very precious tumour sections which were accidentally stained with the wrong primary antibody followed by DAB. They want to know if it is possible to destain the sections (remove the DAB, which is, apparently, quite dark). They have experience removing immunofluorescence antibody staining but not the DAB product.
> All suggests will be very gratefully accepted.
> Many thanks
> Paul
>
> Assoc. Prof. Paul Rigby
> Optical Microscopy Specialist
> Centre for Microscopy, Characterisation & Analysis (M510)
> The University of Western Australia
> 35 Stirling Highway
> Crawley  WA  6007
> Australia

[Francisco J H Blazquez]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hello
Can't they use another (the correct) primary antibody and use another
chromogen? Like a double staining?
Or redo it using immunofluorescence? In this case the DAB will not be seen.

On 01/04/2015 02:08, Paul Rigby wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi All,
> One of our users has had a mishap with some very precious tumour sections which were accidentally stained with the wrong primary antibody followed by DAB. They want to know if it is possible to destain the sections (remove the DAB, which is, apparently, quite dark). They have experience removing immunofluorescence antibody staining but not the DAB product.
> All suggests will be very gratefully accepted.
> Many thanks
> Paul
>
> Assoc. Prof. Paul Rigby
> Optical Microscopy Specialist
> Centre for Microscopy, Characterisation & Analysis (M510)
> The University of Western Australia
> 35 Stirling Highway
> Crawley  WA  6007
> Australia
>

--
Prof. Dr. Francisco Javier Hernandez Blazquez
University of São Paulo

[Sathya]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Paul,
Just to add another discussion topic in IHC World about DAB staining and de-staining:
 
http://www.ihcworld.com/smf/index.php?topic=1458.0
 
Hope this helps and good luck.
 
Sathya Srinivasan
Manager
RUN advanced optical microscopy core facility
(www.ucalgary.ca/runcore)
University of Calgary
Calgary, AB
 

> Date: Wed, 1 Apr 2015 13:54:09 +0000
> From: [hidden email]
> Subject: Re: Removing DAB IHC staining
> To: [hidden email]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I found this post where apparently someone managed to accidentally destain their DAB stained tissues.  Sounds like a bit of triton-X at 4oC over a weekend does the trick: http://www.ihcworld.com/smf/index.php?topic=666.0
>
> -Ben Smith
>
> Benjamin E. Smith, Ph.D.
> Samuel Roberts Noble Microscopy Laboratory
> Research Scientist, Confocal Facility Manager
> University of Oklahoma
> Norman, OK 73019
> E-mail: [hidden email]
> Voice   405-325-4391
> FAX  [phone]
> http://www.microscopy.ou.edu/
> ________________________________________
> From: Confocal Microscopy List [[hidden email]] on behalf of Michael Schell [[hidden email]]
> Sent: Wednesday, April 01, 2015 8:47 AM
> To: [hidden email]
> Subject: Re: Removing DAB IHC staining
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> The only thing I know that works for this is fish.  Some breeds of tiny fish find the DAB precipitate to be quite the delicacy.  Fill your Coplin jar with the purest triple-distilled water, then add fish.  You need the type used in toe salons.  It will take a while, so be sure to change the fish frequently.  The fish must be very hungry for this to work.
>
>
> > On Apr 1, 2015, at 1:08 AM, Paul Rigby <[hidden email]> wrote:
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your posting.
> > *****
> >
> > Hi All,
> > One of our users has had a mishap with some very precious tumour sections which were accidentally stained with the wrong primary antibody followed by DAB. They want to know if it is possible to destain the sections (remove the DAB, which is, apparently, quite dark). They have experience removing immunofluorescence antibody staining but not the DAB product.
> > All suggests will be very gratefully accepted.
> > Many thanks
> > Paul
> >
> > Assoc. Prof. Paul Rigby
> > Optical Microscopy Specialist
> > Centre for Microscopy, Characterisation & Analysis (M510)
> > The University of Western Australia
> > 35 Stirling Highway
> > Crawley  WA  6007
> > Australia