Application
Immunohistochemical staining of bNOS in rat brain using
Application
Immunohistochemistry was performed on normal biopsies of deparaffinized mouse skeletal muscle tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, 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 1:400 with a Rabbit Polyclonal Antibody recognizing nNOS 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.
Application
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 (pH 6.0) buffer, 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 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.
Application
Western blot analysis of nNOS was performed by loading 40 µg 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 recognizing nNOS at a dilution of 1:1000.