[Histonet] SMC Immuno and Elastin combo?

Thomas, Nancy nto <@t> stowers.org
Wed Mar 13 11:32:54 CDT 2013


Patti,
The immunostaining probably was not wiped out, rather just covered up with the red color of the VG counterstain.  You might try a chromogen that expresses in a different color.  Or, you might leave off the van gieson part of the EVG (staining only the elastic tissue) and pick a counterstain of another color.  Basically, you just need to make sure that none of the stains used have the same color outcome.  I have done this combo in the past.  Never really finished the project because the issue I had was that the lizard tissue I was working with just would not stay on the slide no matter what I tried.  But, I do remember that I treated them as two separate stains.   I did the actin staining first, rinsed it well, then started in on the elastin stain.  You probably have a protocol for each of these and you just need to combine them. 

Nancy Thomas	
Stowers Institute

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patricia Bourassa
Sent: Wednesday, March 13, 2013 10:58 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] SMC Immuno and Elastin combo?

Hello,

I've read some papers which reference the combination of smooth muscle cell actin immunostaining with a Verhoeff's elastin counterstain.

Has anyone ever done this and if so, do you have protocol you'd be willing to share?

I'm staining mouse and rat lung and have a SMC protocol working that uses
an Abcam antibody and NovaRed chromagen (peroxidase).   I've also done EVG
staining so I'm familiar with the general process.  The combination is what I'm not sure of - I gave it a try for the heck of it and wiped out the immunostain.

We've just started looking at this, so nothing is set in stone and I'd be grateful for any advice/ procedural help.

Thanks in advance!

Patti Bourassa
Karos Pharmaceuticals
--
*Patti Bourassa
Senior Scientist
Karos Pharmaceuticals
(203) 535-0540, ext 207*
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