[Histonet] Integrin beta1 / CD29 -- cell surface or not??

From:GT Hebert

Hello all,
Does anyone have experience with the antigen?  I am using a monoclonal rat anti-mouse integrin beta1 (clone 265917) from R&D systems with a goat anti-rat (Alexa-594 conj.) secondary -- DAPI nuclear staining.   I am staining mouse aortic sinuses with atherosclerotic lesions.  Today was my first run and I expected to get nice cell membrane staining red.  However, I am getting little 'cell' membrane and more nuclear membrane (?) staining.  How can this be?  The specificity should be to "detect mouse Integrin beta1" -- CD29 (cell surface??).  My negatives are mostly clear except for some non-specific binding (Ab concentration to high) along the smooth muscle cells -- no nuclear staining seen in negative tells me my positive is true positive.
I need a really good cell surface marker so that I can run double immunos to determine if there is co-localization of my antigen on the cell surface.  If anyone has any suggestions, answers or questions for that matter ... anything would be very helpful.  Also, if you have run immunofluorescence with the antibody or other well-definied cell surface markers, photos of what I am supposed to be seeing would also be very helpful.
Thank you.
Gustave Hebert
Scientist II
Wyeth Research
Cambridge MA
email photos: emerald_lake77@yahoo.com

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