Western blot analysis of nNOS was performed by loading 40µg of Mouse (Lane 1) and Rat Brain (Lane 2) tissue lysate onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a Nitrocellulose membrane. Membranes were probed with a rabbit polyclonal antibody (ab3511) recognizing nNOS at a dilution of 1:1000.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse skeletal muscle tissue sections labeling nNOS (neuronal) with ab3511 at 1/400 (left). Antigen retrieval was performed using 10M sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Tissues were blocked in 3% BSA in PBS for 30 minutes at room temperature. A biotin-conjugated secondary antibody was used. Right - negative control.

Immunohistochemistry was performed on normal biopsies of deparaffinized Rat cerebellum tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer and microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a Rabbit Polyclonal Antibody recognizing nNOS (ab3511 ) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

ab3511 at 1/800 dilution staining nNOS (neuronal) in rat brain tissue section by immunohistochemistry (Frozen sections).