[Histonet] Alpha Smooth Muscle Actin -IHC

[fmoraes <@t> igc.gulbenkian.pt]

Hi Wulan Anggrahini

I am not working with cryostat sections only with Whole Mount E10.5 mouse
embryos. I bought the Cy3 conjugated mouse monoclonal anti-Smooth Muscle Actin
(clone 1A4 SIGMA C6198) and once is conjugated with Cy3 you don't need a 2ary
Antibody. It works very well with a 1:400 dilution.
Hope it helps

Filipa

--------------------------------
Filipa Moraes
Moisés Mallo's Lab
Neural Crest Group
Instituto Gulbenkian de Ciência
Rua da Quinta Grande 6
2800-156, Oeiras
Portugal
URL: www.igc.gulbenkian.pt
fmoraes <@t> igc.gulbenkian.pt
Phone: +351 214464525





Message: 10
> Date: Wed, 12 Jul 2006 01:26:15 -0400
> From: "Andrea T. Hooper" <anh2006 <@t> med.cornell.edu>
> Subject: Re: [Histonet] Alpha Smooth Muscle Actin -IHC
> To: Wulan Anggrahini <wulan <@t> med.kobe-u.ac.jp>,	Histonet
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <p06200702c0da377754b4@[140.251.51.71]>
> Content-Type: text/plain; charset="us-ascii" ; format="flowed"
>
> Use DAKO's EPOS version. It's pricey but works incredibly well with
> ZERO background on mouse tissue.  Although I am not familiar with the
> secondary reagents you used it is likely that the background is from
> the anti-mouse secondary reacting with endogenous IgG in the tissue.
> If you cannot get access to EPOS you must use a mouse-on-mouse
> detection system. Contact me privately if you need more information.
>
> Hope this helps!
> Andrea
>
>
>
> >Is there anyone doing IHC for alpha SMA (mouse anti-aSMA Sigma,
> >clone 1A4) on mouse cryostat section ? I did on carotid artery
> >tissue and I used 1/100 dilution overnight and secondary antibody is
> >Histofine Max-PO (Mouse) in 1/200 dilution overnight.
> >
> >Problem is keep having background with low signal/background ratio.
> >Anyone has any tips ? I keep trying for 2 months already and now it
> >starts to make me frustating. Thanks before
> >
> >Wulan Anggrahini
> >Japan
> >
>
> --
>