Mouse monoclonal anti gaba antibody

The animals (n = 30) were deeply anesthetized through immersion in carbonate-buffered tap water containing MS-222 (100 mg/l; Sigma, St Louis, MO, USA). Following decapitation, a portion of the spinal cord, rostral to the dorsal fin, was fixed by 4% (wt/vol) paraformaldehyde (PFA) in phosphate-buffered saline (PBS) for 12–24 hr at 4°C, and subsequently cryoprotected in 20% sucrose in phosphate buffer (PB) for 3–12 hr. For GABA and dopamine immunodetection, 1% glutaraldehyde (vol/vol) was added to the fixative solution. Transverse sections (20 µm thick) were cut on a cryostat (Microm HM 560, Microm International GmbH, Walldorf, Germany), collected on gelatine-coated slides, and kept at −20°C until processing. Sections were incubated/co-incubated with different primary antibodies listed here: a mouse monoclonal anti-acetylated tubulin antibody (dilution 1:500, Sigma-Aldrich), a rat monoclonal anti-somatostatin antibody (1:100, Millipore), a rabbit polyclonal anti-somatostatin-14 IgG antibody (1:1000, Peninsula laboratories), a mouse monoclonal anti-TH antibody (1:200, Millipore), a rabbit polyclonal anti-TH antibody (1:500, Millipore), a mouse monoclonal anti-dopamine antibody (1:400, Millipore), and a mouse monoclonal anti-GABA antibody (1:2000, Swant).