Human/Mouse/Rat nNOS Antibody

  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat nNOS in Western blots. In direct ELISAs, this antibody shows approximately 5% cross‑reactivity with recombinant human eNOS.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human nNOS
    Ser218-Ser1434
    Accession # P29475
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunohistochemistry
    1-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse and Rat nNOS by Western Blot. Western blot shows lysates of mouse brain tissue and rat brain tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse/Rat nNOS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2416) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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.
Immunohistochemistry
nNOS in Human Brain. nNOS was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using 1.7 µg/mL Goat Anti-Human/Mouse/Rat nNOS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2416) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm of astrocytes in the cortex. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
nNOS in Rat Brain. nNOS was detected in immersion fixed frozen sections of rat brain (cortex) using Goat Anti-Human/Mouse/Rat nNOS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2416) at 5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to neurons and neuronal processes. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Mouse and Rat nNOS by Simple WesternTM. Simple Western lane view shows lysates of mouse brain tissue and rat brain tissue, loaded at 0.2 mg/mL. A specific band was detected for nNOS at approximately 160 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat nNOS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2416) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: nNOS
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.
  • Long Name:
    Neuronal Nitric Oxide Synthase
  • Entrez Gene IDs:
    4842 (Human); 18125 (Mouse)
  • Alternate Names:
    bNOS; brain; Constitutive NOS; EC 1.14.13.39; IHPS1; Neuronal NOS; nitric oxide synthase 1 (neuronal); nNOS; N-NOS; nNOSNOS type I; NOS; NOS1; Peptidyl-cysteine S-nitrosylase NOS1
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