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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine

Cited:

Human, Mouse, Rat, Porcine

Applications

Validated:

Flow Cytometry, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, In vitro assay, Western Blot

Cited:

Block/Neutralize, Electron Microscopy, Flow Cytometry, IF/IHC, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

0.5 mg/ml

Product Specifications

Immunogen

Sythetic peptide made to an internal portion of mouse iNOS (between amino acids 12-48) [UniProt P29477].

Reactivity Notes

Porcine reactivity reported in scientific literature (PMID: 31292486).

Specificity

This antibody detects iNOS. It does not detect other NOS isoforms.

Clonality

Polyclonal

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.

Scientific Data Images for iNOS Antibody - BSA Free

Western Blot: iNOS AntibodyBSA Free [NB300-605]

Western Blot: iNOS AntibodyBSA Free [NB300-605]

Western Blot: iNOS Antibody [NB300-605] - Analysis of iNOS was performed by loading 20 ug of RAW264 whole cell lysate untreated (left lane) or stimulated with LPS at 1 ug/mL for 16 hours (right lane) and 10 uL of PageRuler Plus Prestained Protein Ladder onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% Milk in TBST for at least 1 hour. The membrane was probed with an iNOS Rabbit polyclonal antibody at a dilution of 1:1000 overnight at 4C on a rocking platform, washed in TBST, and probed with a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at a dilution of 1:1000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico.
Immunohistochemistry-Paraffin: iNOS Antibody - BSA Free [NB300-605]

Immunohistochemistry-Paraffin: iNOS Antibody - BSA Free [NB300-605]

Immunohistochemistry-Paraffin: iNOS Antibody [NB300-605] - Immunohistochemistry was performed on normal deparaffinized human Lung tissue.
Immunohistochemistry-Paraffin: iNOS Antibody - BSA Free [NB300-605]

Immunohistochemistry-Paraffin: iNOS Antibody - BSA Free [NB300-605]

Immunohistochemistry-Paraffin: iNOS Antibody [NB300-605] - Immunohistochemistry was performed on normal deparaffinized human Heart tissue.

Applications for iNOS Antibody - BSA Free

Application
Recommended Usage

Flow Cytometry

reported in scientific literature (PMID 31536479)

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Frozen

reported in scientific literature (PMID 35005642)

Immunohistochemistry-Paraffin

1:20

In vitro assay

reported in scientific literature (PMID 27998907)

Western Blot

1:200 - 1:800
Application Notes
WB: Detects an approx. 135 kDa protein representing recombinant human iNOS and human iNOS from cytokine stimulated A549 cells. Also detects purified recombinant mouse iNOS, mouse iNOS from cytokine stimulated RAW 264.7 cells and cytokine stimulated rat fibroblast iNOS. However, the signals are not as strong as those seen with the human samples.

Reviewed Applications

Read 3 reviews rated 4.7 using NB300-605 in the following applications:

Formulation, Preparation, and Storage

Purification

Affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: iNOS

Nitric oxide (NO) is a colorless, free radical gas that carries a variety of messages between cells. Vasorelaxation, neurotransmission and cytotoxicity can all be potentiated through cellular response to NO. NO production is mediated by members of the nitric oxide synthase (NOS) family including the two constitutive isoforms: brain, bNOS, or neuronal NOS, nNOS (type I) and endothelial cell NOS, eNOS (type III); along with the inducible isoform, iNOS (type II). NOS catalyzes the oxidization of L-arginine to produce L-citrulline and NO, requiring the cofactors calmodulin, nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), and flavin mononucleotide (FMN), heme, and tetrahydrobiopterin (1).

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

Entrez Gene IDs

4843 (Human); 18126 (Mouse); 24599 (Rat)

Gene Symbol

NOS2

UniProt

Additional iNOS Products

Product Documents for iNOS Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for iNOS Antibody - BSA Free

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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