RE: direct peroxidase for CD31
Without knowing the particulars of the experiment, your post-doc is choosing
the LEAST sensitive technique. Direct IHC is more commonly used for
immunofluorescence in my experience, but even then most researchers use at
least a 2-step method. I use anti-murine CD31 and I don't even believe it is
available commercially as an HRP conjugate, thus as Patsy said, you would
have to do that yourself and verify the labeling efficacy each time you
do(so much for speed).
At the very least you can get biotinylated anti-mouse CD31 and go from
Those who are serious about getting more complete data out of an experiment
generally opt the MOST sensitive technique. Many have used ABC or LSAB for
CD31 detection in mice without the problems he mentions. I think he needs to
be more open-minded, it will serve him well in the future.
Brett M. Connolly, Ph.D.
Merck Research Laboratories
Dept. of Pharmacology
PO Box 4
West Point, PA 19486
From: Thu-Thao Pham [mailto:firstname.lastname@example.org]
Sent: Tuesday, November 20, 2001 5:42 PM
To: DAKOTechServ@dakousa.com; email@example.com
Subject: direct peroxidase for CD31
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.
>Subject: RE: DAKO kit for CD31
>Date: Tue, 20 Nov 2001 13:28:53 -0800
>First, I would like to say "Thank you" to Bob Meyer and Patsy Ruegg for
>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.
>Technical Service Specialist
>From: firstname.lastname@example.org [mailto:email@example.com]
>Sent: Tuesday, November 20, 2001 10:56 AM
>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
>a labelled streptavidin-biotin system for use with primary antibodies
>in mouse or rabbit, but it is optimized for use with DAKO N-Series ready to
>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
>a lot of their products.
>Bob Meyer, HTL
> > I looked through the catolog and don't understand the difference between
> > catolog:
> > cat. K0681
> > Universal DAKO LSAB (labelled Streptavind-Biotin) system, Peroxidase for
> > with rabbit or mouse antibody.
> > cat. K0675
> > Universal DAKO LSAB2 (labelled Streptavind-Biotin) system, Peroxidase
> > use with rabbit or mouse antibody.
> > Thao
> > _________________________________________________________________
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