AB5382 | Anti-Nitric Oxide Synthase II Antibody

Western Blot Analysis:
Representative lot data.
Mouse liver lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with anti-INOS (1:1000 dilution). Proteins were visualized using a donkey anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates protein INOS (~131 kDa)
Western Blot Analysis:
Representative lot data.
Mouse liver lysate was resolved by electro
AB5382
50 µL  
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      Overview

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      R, MWB, ICC, IHCRbSerumPolyclonal Antibody
      Description
      Catalogue Number AB5382
      Brand Family Chemicon®
      Synonyms iNOS NOS II
      Trade Name
      • Chemicon
      Description Anti-Nitric Oxide Synthase II Antibody
      Background Information Nitric Oxide Synthases are the only enzymes know to simultaneously require five bound cofactors, FAD, FMN, haem, tetrahydrobiopterin (BH4) and Ca++-Calmodulin (CAM). In mammals, there are three distinct genes for NOS, NOS I is neuronal, NOS II is cytokine inducible, and NOS III is endothelial. The enzymes exist as homodimers, each monomer consisting of two major domains: N­terminal oxygenase domain, which belongs to the class of haem-thiolate proteins, and C­terminal reductase domain, which is homologous to NADPH:P450 reductase. The interdomain linker between the oxygenase and reductase domains contains a CaM­binding sequence. Functionally NOS is an enzyme that catalyzes the stepwise conversion of the amino acid L-arginine to nitric oxide and L-citrulline. NOS activity stimulates cyclic GMP, inducing smooth muscle relaxation by increasing intracellular cGMP, which in turn inhibits calcium entry into the cell, and decreases intracellular calcium concentrations. This also stimulates a cGMP-dependent protein kinase that activates myosin light chain phosphatase, the enzyme that dephosphorylates myosin light chains, which leads to smooth muscle relaxation.
      References
      Product Information
      Format Serum
      Specificity Inducible nitric oxide synthase (iNOS, NOS-II). No cross-reaction with rat nNOS and eNOS by Western blot and immunohistochemistry.
      Control
      • Macrophages, liver lysates
      Presentation Rabbit polyclonal serum in buffer containing liquid 0.05% sodium azide.
      Applications
      Application Anti-Nitric Oxide Synthase II Antibody is an antibody against Nitric Oxide Synthase II for use in IC, IH & WB.
      Key Applications
      • Western Blotting
      • Immunocytochemistry
      • Immunohistochemistry
      Application Notes Immunohistochemistry:
      1:5000 (ABC) dilution of a previous lot was used on tissue sections (rat liver).

      Immunocytochemistry:
      1:50,000 dilution of a previous lot was used on cultured cells.

      Western blot:
      1:5,000 (ECL). The antibody has been used successfully on rat liver. It is not recommended for use on rat brain extracts since unidentified bands may be present.

      Optimal working dilutions must be determined by end user.
      Biological Information
      Immunogen species C-terminal 14 amino acid peptide from mouse macrophage nitric oxide synthase.
      Epitope C-terminus
      Concentration Please refer to the Certificate of Analysis for the lot-specific concentration.
      Host Rabbit
      Specificity Inducible nitric oxide synthase (iNOS, NOS-II). No cross-reaction with rat nNOS and eNOS by Western blot and immunohistochemistry.
      Species Reactivity Rat Mouse
      Species Reactivity Note Mouse and rat. Reactivity to human iNOS is approximately 1%.
      Antibody Type Polyclonal Antibody
      Entrez Gene Number
      Entrez Gene Summary Nitric oxide is a reactive free radical which acts as a biologic mediator in several processes, including neurotransmission and antimicrobial and antitumoral activities. This gene encodes a nitric oxide synthase which is expressed in liver and is inducible by a combination of lipopolysaccharide and certain cytokines. Three related pseudogenes are located within the Smith-Magenis syndrome region on chromosome 17.
      Gene Symbol
      • EC 1.14.13.39
      • NOS2A
      • iNOS
      • HEP-NOS
      • INOS
      • NOS
      • NOS2
      Purification Method Unpurified
      UniProt Number
      UniProt Summary FUNCTION: SwissProt: P35228 # Produces nitric oxide (NO) which is a messenger molecule with diverse functions throughout the body. In macrophages, NO mediates tumoricidal and bactericidal actions.
      COFACTOR: Heme group (By similarity). & Binds 1 FAD (By similarity). & Binds 1 FMN (By similarity). & Metrahydrobiopterin (BH4). May stabilize the dimeric form of the enzyme (By similarity).
      SIZE: 1153 amino acids; 131117 Da
      SUBUNIT: Homodimer. Binds SLC9A3R1.
      TISSUE SPECIFICITY: Expressed in the liver, retina, bone cells and airway epithelial cells of the lung. Not expressed in the platelets.
      SIMILARITY: SwissProt: P35228 ## Belongs to the NOS family. & Contains 1 flavodoxin-like domain.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality Assurance Evaluated by Western Blot on Mouse liver lysates.

      Western Blot Analysis:
      1:1000 dilution of this antibody detected INOS on 10 µg of Mouse liver lysates.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage Conditions Stable for 6 months at -20°C in undiluted aliquots from date of receipt.
      Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
      Packaging Information
      Material Size 50 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Languages
      English

      Certificates of Analysis

      TitleLot Number
      Anti-Nitric Oxide Synthase II 2459002
      Anti-Nitric Oxide Synthase II - 2392230 2392230
      Anti-Nitric Oxide Synthase II - 2064696 2064696
      Anti-Nitric Oxide Synthase II - 2227120 2227120
      Anti-Nitric Oxide Synthase II - JBC1766784 JBC1766784
      Anti-Nitric Oxide Synthase II - JC1650285 JC1650285
      Anti-Nitric Oxide Synthase II - JC1690726 JC1690726
      Anti-Nitric Oxide Synthase II - LV1501936 LV1501936
      Anti-Nitric Oxide Synthase II - NG1723923 NG1723923
      Anti-Nitric Oxide Synthase II - NG1798879 NG1798879
      Anti-Nitric Oxide Synthase II - NG1873244 NG1873244
      Anti-Nitric Oxide Synthase II -2502912 2502912

      References

      Reference overviewPub Med ID
      Proteomic characterization of the cellular response to nitrosative stress mediated by s-nitrosoglutathione reductase inhibition.
      Matthew W Foster,Zhonghui Yang,David M Gooden,J Will Thompson,Carol H Ball,Meredith E Turner,Yongyong Hou,Jingbo Pi,M Arthur Moseley,Loretta G Que
      Journal of proteome research 11 2012

      Show Abstract
      22390303
      Dexamethasone inhibits the nox-dependent rOS production via suppression of MKP-1-dependent MAPK pathways in activated Microglia.
      Huo Y, Rangarajan P, Ling EA, Dheen ST
      BMC neuroscience 12 49. 2011

      Show Abstract Full Text Article
      21615929
      Peripheral purinergic receptor modulation influences the trigeminal ganglia nitroxidergic system in an experimental murine model of inflammatory orofacial pain.
      Borsani, Elisa, et al.
      J. Neurosci. Res., 88: 2715-26 (2010) 2010

      Show Abstract
      20648657
      Distinct pattern of microglial response, cyclooxygenase-2, and inducible nitric oxide synthase expression in the aged rat brain after excitotoxic damage.
      O Campuzano,M M Castillo-Ruiz,L Acarin,B Castellano,B Gonzalez
      Journal of neuroscience research 86 2008

      Show Abstract
      18543338
      Hepatocyte survival in acute hepatitis is due to c-Jun/AP-1-dependent expression of inducible nitric oxide synthase.
      Peter Hasselblatt, Martina Rath, Vukoslav Komnenovic, Kurt Zatloukal, Erwin F Wagner
      Proceedings of the National Academy of Sciences of the United States of America 104 17105-10 2007

      Show Abstract Full Text Article
      17940019
      Retinoic acid inhibits expression of TNF-alpha and iNOS in activated rat microglia.
      S Thameem Dheen, Yan Jun, Zhou Yan, Samuel S W Tay, Eng Ang Ling
      Glia 50 21-31 2005

      Show Abstract
      15602748
      Astroglial nitration after postnatal excitotoxic damage: correlation with nitric oxide sources, cytoskeletal, apoptotic and antioxidant proteins.
      Laia Acarin,Hugo Peluffo,Luis Barbeito,Bernardo Castellano,Berta González
      Journal of neurotrauma 22 2005

      Show Abstract
      15665612
      Transplanted olfactory ensheathing cells modulate the inflammatory response in the injured spinal cord.
      Rubèn López-Vales, Guillermo García-Alías, Joaquim Forés, José M Vela, Xavier Navarro, Enrique Verdú
      Neuron glia biology 1 201-9 2004

      18634597
      Therapeutic effect of the endogenous fatty acid amide, palmitoylethanolamide, in rat acute inflammation: inhibition of nitric oxide and cyclo-oxygenase systems.
      Costa, Barbara, et al.
      Br. J. Pharmacol., 137: 413-20 (2002) 2002

      Show Abstract
      12359622
      LPS/IFN-gamma cytotoxicity in oligodendroglial cells: role of nitric oxide and protection by the anti-inflammatory cytokine IL-10.
      Molina-Holgado, E, et al.
      Eur. J. Neurosci., 13: 493-502 (2001) 2001

      Show Abstract
      11168556

      Data Sheet

      Title
      Anti-Nitric Oxide Synthase II - Data Sheet