Human/Mouse/Rat iNOS Antibody

(7 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human iNOS. By using synthetic peptides, the epitope recognized by this antibody has been mapped to aa 781-798 of human iNOS. The corresponding sequence of mouse iNOS is identical.
  • Source
    Monoclonal Mouse IgG1 Clone # 2D2-B2
  • Purification
    Protein A or G purified from ascites
  • Immunogen
    Recombinant human iNOS
    Pro781-His798
    Accession # P35228
  • 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
    8-25 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 Human iNOS by Western Blot. Western blot shows lysates of DLD clone 2C2 human colon adenocarcinoma cell line untreated (-) or treated (+) with Recombinant Human IL-1 beta, Recombinant Human TNF-alpha, and Recombinant Human IFN-gamma (Catalog #201‑LB,
210‑TA,
285‑IF) for 24 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat iNOS Monoclonal Antibody (Catalog # MAB9502), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for iNOS at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 6.
Detection of Mouse and Rat iNOS by Western Blot. Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line and NR8383 rat alveolar macrophage cell line untreated (-) or treated (+) with 10 μg/mL LPS for 4 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat iNOS Monoclonal Antibody (Catalog # MAB9502) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for iNOS at approximately 125 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
iNOS in Human Brain. iNOS was detected in immersion fixed paraffin-embedded sections of human brain (medulla) using Mouse Anti-Human/Mouse/Rat iNOS Monoclonal Antibody (Catalog # MAB9502) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Mouse iNOS by Simple WesternTM. Simple Western lane view shows lysates of RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with LPS, loaded at 0.2 mg/mL. A specific band was detected for iNOS at approximately 136 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse/Rat iNOS Monoclonal Antibody (Catalog # MAB9502). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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: iNOS
Inducible nitric oxide synthase (iNOS) is a widely expressed, approximately 130 kDa enzyme that catalyzes the conversion of L-arginine to nitric oxide (NO) plus citrulline. NO is a bioactive mediator that plays an important role in hemodynamics by inducing vascular smooth muscle relaxation. iNOS is constitutively expressed in the kidney. In other tissues, it is inducibly expressed during inflammation, oxidative stress, and hyperglycemia. iNOS activity is elevated in a variety of disease states including atherosclerosis, heart failure, sepsis, solid tumors, and type 2 diabetes. Increased production of NO mediates acute kidney injury, TNF-alpha induced muscle wasting, and the increased radiosensitivity of hypoxic tumor cells. Within amino acids 781-798, human iNOS shares 89% and 72% aa sequence identity with mouse and rat iNOS, respectively.
  • Long Name:
    Inducible Nitic Oxide Synthase
  • Entrez Gene IDs:
    4843 (Human); 18126 (Mouse)
  • Alternate Names:
    EC 1.14.13.39; Hepatocyte NOS; HEP-NOSPeptidyl-cysteine S-nitrosylase NOS2; Inducible NO synthase; Inducible NOS; iNOS; nitric oxide synthase 2, inducible; nitric oxide synthase 2A (inducible, hepatocytes); nitric oxide synthase, inducible; nitric oxide synthase, macrophage; NOS; NOS, type II; NOS2; NOS2A; NOS2ANOS type II
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
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Species
Applications
Sample Type
  1. Erythropoietin Receptor Antagonist Suppressed Ectopic Hemoglobin Synthesis in Xenografts of HeLa Cells to Promote Their Destruction.
    Authors: Yasuda Y, Fujita M, Koike E, Obata K, Shiota M, Kotani Y, Musha T, Tsuji-Kawahara S, Satou T, Masuda S, Okano J, Yamasaki H, Okumoto K, Uesugi T, Nakao S, Hoshiai H, Mandai M
    PLoS ONE, 2015;10(4):e0122458.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  2. High pre-transplant serum nitrate levels predict risk of acute steroid-refractory graft-versus-host disease in the absence of statin therapy.
    Authors: Dietrich S, Okun J, Schmidt K, Falk C, Wagner A, Karamustafa S, Radujkovic A, Hegenbart U, Ho A, Dreger P, Luft T
    Haematologica, 2014;99(3):541-7.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Artichoke, cynarin and cyanidin downregulate the expression of inducible nitric oxide synthase in human coronary smooth muscle cells.
    Authors: Xia N, Pautz A, Wollscheid U, Reifenberg G, Forstermann U, Li H
    Molecules, 2014;19(3):3654-68.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Dose-dependent changes in neuroinflammatory and arachidonic acid cascade markers with synaptic marker loss in rat lipopolysaccharide infusion model of neuroinflammation.
    BMC Neurosci, 2012;13(0):50.
    Species: Rat
    Sample Type: Tissue Homogenates
    Application: WB
  5. The autoantigen Ro52 is an E3 ligase resident in the cytoplasm but enters the nucleus upon cellular exposure to nitric oxide.
    Authors: Espinosa A, Oke V, Elfving A, Nyberg F, Covacu R, Wahren-Herlenius M
    Exp. Cell Res., 2008;314(20):3605-13.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  6. Proinflammatory and antimicrobial nitric oxide in gingival fluid of diabetic patients with periodontal disease.
    Authors: Skaleric U, Gaspirc B, McCartney-Francis N, Masera A, Wahl SM
    Infect. Immun., 2006;74(12):7010-3.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  7. Attenuation of apoptosis and enhancement of proteoglycan synthesis in rabbit cartilage defects by hyperbaric oxygen treatment are related to the suppression of nitric oxide production.
    Authors: Yuan LJ, Ueng SW, Lin SS, Yeh WL, Yang CY, Lin PY
    J. Orthop. Res., 2004;22(5):1126-34.
    Species: Rabbit
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
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