I used this antibody on PFA fixed, sucrose cryoprotected frozen sections from the mouse brain (below is a brief description). The primary a.b. I used was anti-mGluR5 (ab53090), which was somewhat difficult to get working.
Transcardially perfused the mouse with 4% PFA. Dissected the brain and placed in PFA for a few hours later, placed in 30% sucrose overnight for cryoprotection. Then, snap/flash froze the brain and used a cryostat for sectioning into 20 micron coronal slices. Applied primary anti-mGluR5 (ab53090) antibody, followed by the secondary antibody (dilution: 1/200), according to standard protocols. No blocking, no antigen-retrieval, and no permeabilization was done.
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Submitted Jan 29 2014