iNOS Antibody (PSH03-69)
Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32473
Recombinant Monoclonal Antibody
Key Product Details
Species Reactivity
Human, Mouse
Applications
Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # PSH03-69
Concentration
1 mg/ml
Product Specifications
Immunogen
Synthetic peptide. (Uniprot: P35228)
Localization
Cytoskeleton, Nucleus, Cytosol.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
131 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for iNOS Antibody (PSH03-69)
Western Blot: iNOS Antibody (PSH03-69) [NBP3-32473]
Western Blot: iNOS Antibody (PSH03-69) [NBP3-32473] - Western blot analysis of iNOS on different lysates with Rabbit anti-iNOS antibody (NBP3-32473) at 1/1,000 dilution.Lane 1: RAW264.7 cell lysate
Lane 2: RAW264.7 treated with 1ug/mL LPS for 24 hours cell lysate
Lysates/proteins at 10 ug/Lane.
Predicted band size: 131 kDa
Observed band size: 131 kDa
Exposure time: 3 minutes; ECL;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32473) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemistry: iNOS Antibody (PSH03-69) [NBP3-32473]
Immunohistochemistry: iNOS Antibody (PSH03-69) [NBP3-32473] - Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-iNOS antibody (NBP3-32473) at 1/200 dilution.The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32473) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunocytochemistry/ Immunofluorescence: iNOS Antibody (PSH03-69) [NBP3-32473]
Immunocytochemistry/ Immunofluorescence: iNOS Antibody (PSH03-69) [NBP3-32473] - Immunocytochemistry analysis of RAW264.7 cells treated with or without 1μg/mL LPS for 6 hours labeling iNOS with Rabbit anti-iNOS antibody (NBP3-32473) at 1/100 dilution.Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-iNOS antibody (NBP3-32473) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
Applications for iNOS Antibody (PSH03-69)
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
1:100
Immunohistochemistry-Paraffin
1:200
Western Blot
1:1000
Formulation, Preparation, and Storage
Purification
Protein A purified
Formulation
PBS (pH7.4), 0.1% BSA and 40% Glycerol
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Background: iNOS
The 131 kDa enzyme, iNOS, is found in a variety of cell types including macrophages, hepatocytes, synoviocytes, and smooth muscle cells. While constitutively expressed in kidneys, in other tissues iNOS is induced by bacterial lipopolysaccharides (LPS), growth factors, and cytokines such as IFN-gamma, TNF, IL-1 and IL-2. iNOS is not regulated by the level of intracellular Ca2+ and is constantly active as a dimer when expressed. iNOS activity is elevated in a variety of diseases including atherosclerosis, heart failure, sepsis, solid tumors, and type 2 diabetes. Acting as a critical mediator of inflammation and apoptosis, iNOS inhibitors have been shown to alleviate obesity and stress inducted insulin resistance in mouse models (2,3).
References
1. Forstermann U, and Sessa WC. (2012) Nitric oxide synthases: regulation and function. Eur Heart J. 33(7): 829-837. PMID: 21890489
2. Aktan F. (2004) iNOS-mediated nitric oxide production and its regulation. Life Sci. 75(6):639-53. PMID: 15172174
3. Cinelli MA, Do HT, Miley GP, Silverman RB. (2020) Inducible nitric oxide synthase: Regulation, structure, and inhibition. Med Res Rev. 40(1):158-189. PMID: 31192483
Long Name
Inducible Nitic Oxide Synthase
Alternate Names
NOS2, NOS2A
Gene Symbol
NOS2
Additional iNOS Products
Product Documents for iNOS Antibody (PSH03-69)
Product Specific Notices for iNOS Antibody (PSH03-69)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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