[Histonet] Mouse GranzymeB

[Milne, Katy]

Hi Amos,

We use cat # ab4059 from Abcam for our mouse (and human) granzyme B.

It does have some nuclear background too but when we use it in conjunction with Biocare's Renaissance Background Reducing Diluent it cleans it up nicely.

As you already have the Ab in, you could always try ordering some of the diluent (it's pretty cheap), see if that cleans it up.

Good luck!  Let me know if you have any other questions!  :)

Katy  



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Message: 21
Date: Fri, 24 Jan 2014 12:09:09 -0500
From: Amos Brooks <amosbrooks <@t> gmail.com>
Subject: [Histonet] Mouse GranzymeB
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Ok Musketeers,
     I am trying to detect cytotoxic T-Cells in formalin fixed paraffin
embedded mouse liver. GranzymeB should do the trick. I have a rabbit anti
mouse (and human and rat) GranzymeB from abCam (ab53097). The spleen that I
ran as a control with it works fine. Nice T-cells and no signifigant
background staining. The liver on the others hand has some lymphocyte
staining, presumably cytotoxic T-cells as expected,  but it looks like
every hepatocyte nuclei are also picking it up.  Grrr,  right?
     If this antibody had been raised in mouse,  I wouldn't be surprised.
Also,  if the spleen looked similar, I wouldn't be surprised. Do any of you
have any ideas ab pi ut what this might be? Alternatively, does anyone have
any ideas about another antibody that would work here?  CD4 or CD8 would be
great here,  but its FFPE mouse,  so sadly that's out (more grumbling).
    For the completionists out there,  I used EDTA pH9,  peroxide block and
detected the antibody (diluted 1:800) with Envision (rabbit) and DAB from
Dako.

Any suggestions would be vastly appreciated,

Happy Friday,
Amos


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