Re: direct peroxidase for CD31

From:"C. H."

Thao,

Could the post-doc be meaning for you to use a secondary antibody conjugated to HRP (peroxidase) as opposed to using a biotin amplification indirect method?! 

Cyrla

----Original Message Follows----
From: rueggp
To: Thu-Thao Pham
CC: DAKOTechServ@dakousa.com, histonet@pathology.swmed.edu
Subject: Re: direct peroxidase for CD31
Date: Tue, 20 Nov 2001 18:08:11 -0700
in order to direct peroxidase you will have to find or conjugate the cd31
antibody yourself with peroxidase. does your post-doc know this?
patsy ruegg
Thu-Thao Pham wrote:
> I'm greatful and thankful to everyone that have taken their time to help me
> out with the CD31 method.
>
> I have a problem with a post-doc in the lab that wanted me to do CD31 on a
> mouse skin tissue. However, a tech. before be got fired and the hios lab
> notebook cannot be found. In any case, a post-doc wanted to do a direct
> peroxidase staining, because he said it faster and produce less background
> than using a ABC or LSAB2 method. He don't understand about IHC and he
> cannot have an open minded about modification. I have give this post-doc
> every single information from all of you that response to my questions about
> CD31, but he refused to go through with any method, but "direct peroxidase."
> He said that direct peroxidase method is VERY basic and that everyone should
> know and know how to do it (including myself), but why doesn't he, himself
> tell me how it's suppose to work?
>
> My questions, is does anyone know anything about direct peroxidase and how I
> can approached to do CD31? A post-doc asked me to email around and asked all
> of you to show me how to do direct peroxidase for CD31.
>
> I need to talk to my PI about this Post-doc and the rule in this lab, if I
> responsibility is to do histology for everyone in this lab.
>
> Thao
> >From: DAKOTechServ
> >To: histonet@pathology.swmed.edu
> >Subject: RE: DAKO kit for CD31
> >Date: Tue, 20 Nov 2001 13:28:53 -0800
> >
> >Dear Histonet,
> >
> >First, I would like to say "Thank you" to Bob Meyer and Patsy Ruegg for
> >your
> >replies to this question. Bob, you are correct, the LSAB and LSAB2 systems
> >are designed for mouse and rabbit primaries. The LSAB+ is designed for
> >mouse, rabbit, and goat primaries. The "2" in LSAB2 denotes our second
> >generation of the LSAB system. There were some improvements to the LSAB
> >system that required a second generation status. Please feel free to
> >contact me directly if you would like more information.
> >
> >Sincerely,
> >Danielle Mach
> >Technical Service Specialist
> >DAKO Corporation
> >800-424-0021
> >
> >-----Original Message-----
> >From: r-meyer2@northwestern.edu [mailto:r-meyer2@northwestern.edu]
> >Sent: Tuesday, November 20, 2001 10:56 AM
> >To: histonet@pathology.swmed.edu
> >Subject: Re: DAKO kit for CD31
> >
> >
> >The DAKO LSAB system is for use with primary antibodies produce in mouse or
> >rabbit and is a labelled streptaviding-biotin system. The LSAB2 system is
> >also
> >a labelled streptavidin-biotin system for use with primary antibodies
> >produced
> >in mouse or rabbit, but it is optimized for use with DAKO N-Series ready to
> >use
> >primary antibodies. The LSAB+ system is for use with primary antibodies
> >produced in mouse, rabbit, or goat. And I am not a salesman for DAKO just
> >use
> >a lot of their products.
> >
> >Bob Meyer, HTL
> >Northwestern University
> > >
> > > I looked through the catolog and don't understand the difference between
> >2
> >
> > > catolog:
> > >
> > > cat. K0681
> > > Universal DAKO LSAB (labelled Streptavind-Biotin) system, Peroxidase for
> >use
> > > with rabbit or mouse antibody.
> > >
> > > cat. K0675
> > > Universal DAKO LSAB2 (labelled Streptavind-Biotin) system, Peroxidase
> >for
> > > use with rabbit or mouse antibody.
> > >
> > > Thao
> > >
> > > _________________________________________________________________
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> > >
> > >
> >
> >
>
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