I would guess that there are problems with the antibodies you are using. SMA is usually a very robust antigen for IHC, if you're not getting any staining you're probably using the wrong antibody. You need an antibody whose specs indicate that it works on mouse, and that it works on fixed tissue. You also don't give details on your staining protocol - it could be a technical issue with that as well. To find proper antibodies, I have found the Biocompare website to be an excellent place to look.
Jean-Martin Lapointe AccelLAB Inc ------------------------------ Message: 11 Date: Sat, 7 Mar 2009 00:44:32 +0800 From: "TF" <ti...@foxmail.com> Subject: Re: RE: [Histonet] CD31 on rat brain [Histonet] staining brain vessels > > I was wondering if anybody might have an idea with the following > problem we are experiencing: we want to stain for blood vessels in > sections of mouse brain. Our experimental tissues have been fixed > overnight in 4% paraformaldehyde and have been sitting in PBS since. > We have tried staining with antibodies against desmin, SMA, and > collagen but we get NO specific signal. We recently tried a non-fixed > mouse brain and got desmin to work immediately. The problem is that > we need to use the fixed brains because they are our experimental > model and it would take too long (2 years to be exact) to generate the > same samples. If anybody has come across such a problem before, or > has a specific protocol for vessels that works on PFA fixed brain, we > would appreciate the suggestions! > thanks in advance! > Irini > ----------------------------------------------------- > Irini Skaliora, PhD > Investigator Cá¾½ (Assistant Professor) > Developmental Biology Division > Biomedical Research Foundation of the Academy of Athens (BRFAA) > Soranou Efessiou 4 > Athens 11527 > tel. +30-210-6597203 (office) > tel. +30-210-6597482 (lab) > fax. +30-210-6597545 > email: iskali...@bioacademy.gr _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
