[Histonet] FW: CD86 - mouse

[koellingr <@t> comcast.net]

Joost, 

I was curious myself what was out there and maybe someone has answered you but I haven't seen anything on the HistoNet.  Haven't done anything with murine CD86 (B7.2) for 3-4 years but this is what I can recall.  Of course you know the whole CD80/86/CD28/CTLA-4 costimulatory story is tough because of transient nature and upregulations during stimulation or disease. 



Using the GL-1 clone from BD (don't work for them but like their anti-mouse reagents), we got pretty good results on frozen mouse tissues with standard frozen fixation and IHC. At least reasonable in that staining corresponded to flow data from similar mouse tissues.  As far as FFPE mouse tissues, the best I could do was to get pretty good but not overly convincing data with retieval but with a tyramide amplification necessary.  In flow if you look at a one color diagram of staining (isotype and then GL-1) the peaks are not well separated even with something like LPS stimmed cells. In fact, sometimes the pos peak might move a log but the base of the peaks (isotype and GL-1) can overlap substantially.  On a flow diagram, shoulders and mean peak heights are easy to differentiate over neg or background but I could never really seperate out truely pos from truely negative from low positive in FFPE mouse like you can with flow.  So frozen was the way we went. 



And I know there is, at least in humans, soluble CD86 floating around due to shedding/alternative splicing. If that happens in mice (I'm clueless) that could contribute to background.  GL-1 clone in frozen mouse tissue was how I ended up going but haven't touched this for many years. 



Ray 



Raymond Koelling 

PhenoPath Labs 

Seattle, WA 




----- Original Message ----- 
From: "J.P. Bruijntjes (Joost)" <joost.bruijntjes <@t> tno.nl> 
To: histonet <@t> lists.utsouthwestern.edu 
Sent: Wednesday, March 18, 2009 7:48:00 AM GMT -08:00 US/Canada Pacific 
Subject: [Histonet] FW: CD86 - mouse 

Hi histonetters 

  

Is really nobody working with cd86 on mouse tissues? 

  

Joost Bruijntjes 

TNO Quality of Life 

Zeist 

Holland 

  

  

TNO.NL <http://www.tno.nl/> 

Joost Bruijntjes 

T +31 30 694 44 80 
F +31 30 694 49 86 
E joost.bruijntjes <@t> tno.nl <mailto:joost.bruijntjes <@t> tno.nl> 

Disclaimer <http://www.tno.nl/tno/email/> 

  

This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html 
_______________________________________________ 
Histonet mailing list 
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet