Detection of Human and Mouse nNOS by Western Blot.
Western blot shows lysates of Sf 21 S. frugiperda insect ovarian cell line either mock transfected or transfected with human nNOS, and mouse brain tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse nNOS Monoclonal Antibody (Catalog # MAB24161) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for nNOS at approximately 160 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
nNOS in Neuro‑2A Mouse Cell Line.
nNOS was detected in immersion fixed Neuro‑2A mouse neuroblastoma cell line using Mouse Anti-Human/Mouse nNOS Monoclonal Antibody (Catalog # MAB24161) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei, cytoplasm and cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of nNOS in Neuro‑2A Mouse Cell Line by Flow Cytometry.
Neuro‑2A mouse neuroblastoma cell line was stained with Mouse Anti-Human/Mouse nNOS Monoclonal Antibody (Catalog # MAB24161, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
nNOS is one of three NOS enzymes that catalyze the oxidation of L-argine to L-citruline and nitric oxide. nNOS exists as homodimers containing a cytochrome P450‑like prosthetic heme group in the N-terminal half. It also has a tightly bound FAD and FMN group in the C-terminal half. At least 4 isoforms of human nNOS are known. Human nNOS shares about 55% amino acid sequence identity with eNOS and iNOS. It also shares 96% sequence identity with mouse or rat nNOS.
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