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
>

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