Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine

Cited:

Human, Mouse, Rat, Porcine

Applications

Validated:

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

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all iNOS Primary Antibodies »

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.

Description

Novus Biologicals Rabbit iNOS Antibody - BSA Free (NB300-605) is a polyclonal antibody validated for use in IHC, WB, Flow, ICC/IF and IP. Anti-iNOS Antibody: Cited in 132 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

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.
Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605]

Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605]

Immunocytochemistry/Immunofluorescence: iNOS Antibody [NB300-605] - Analysis of iNOS in A549 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a iNOS polyclonal antibody at a dilution of 1:20 overnight at 4C, washed with PBS and incubated with a DyLight 488 conjugated secondary antibody. iNOS staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown.
Western Blot: iNOS AntibodyBSA Free [NB300-605]

Western Blot: iNOS AntibodyBSA Free [NB300-605]

Western Blot: iNOS Antibody [NB300-605] - iNOS in stimulated astrocytes. Western blot image submitted by a verified customer review.
Western Blot: iNOS AntibodyBSA Free [NB300-605]

Western Blot: iNOS AntibodyBSA Free [NB300-605]

iNOS-Antibody-Western-Blot-NB300-605-img0017.jpg
Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605]

Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605]

Immunocytochemistry/Immunofluorescence: iNOS Antibody [NB300-605] - Analysis of iNOS in NIH-3T3 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a iNOS polyclonal antibody at a dilution of 1:20 overnight at 4C, washed with PBS and incubated with a DyLight 488 conjugated secondary antibody. iNOS staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown.
iNOS Antibody - BSA Free

Western Blot: iNOS Antibody - BSA Free [NB300-605] -

Western Blot: iNOS Antibody - BSA Free [NB300-605] - Stimulation of IMA 2.1 cells had no effect on relative survival after viral infection. IMA 2.1 cells were stimulated with LPS (10 µg/ml), IFN-gamma, (10 ng/ml) LPS + IFN-gamma, IL-4 (10 ng/ml), FGF (5 ng/ml) or FGF + LPS for 24 h in medium with 2% FBS. Polarization of cells was analyzed by Griess assay (a), Western blot (b) & FACS analysis by detection of MHCII+ cells (c). d MTT assay was performed to detect the relative percentage of VACV-mediated cell death in the presence of stimulating factors. e Viral replication was analyzed by standard plaque assay in the presence of stimulating factors. All experiments were performed in triplicate. Image collected & cropped by CiteAb from the following publication (http://www.translational-medicine.com/content/13/1/216), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
iNOS Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605] -

Immunocytochemistry/ Immunofluorescence: iNOS Antibody - BSA Free [NB300-605] - Nfix is mainly expressed by anti-inflammatory MPs. (a) Percentage of F4/80+ MPs positive for Nfix in Tibialis Anterior muscles (TA) of WT mice injected by CTX at D2, D4 & D7, post-injury. Immunostaining for F4/80 (green), Nfix (red) & DAPI (blue) at D4 & D7 after CTX injection; (b) Percentage of Ly6C+ & Ly6C- sorted MPs positive for Nfix in TA muscles of WT mice injected by CTX at D2, D4 & D7 post-injury; (c) Percentage of Nfix+ MPs after M1 & M2c polarization (with IFN gamma & IL10, respectively). * p < 0.05; *** p < 0.001; for (b) * p < 0.05 Ly6C+ vs. Ly6C+ at D4 & D7; # p < 0.05 Ly6C− D7 vs. D2. Results are means ± SEM of at least three independent experiments. Scale bar = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32183151), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
iNOS Antibody - BSA Free

Western Blot: iNOS Antibody - BSA Free [NB300-605] -

Western Blot: iNOS Antibody - BSA Free [NB300-605] - Indolepropionic acid (IPA) induced oxidative stress, cellular energy stress, & decreased the proportions of cancer stem cells. 500,000 cells/well 4T1 cells were treated with IPA in the concentrations indicated for 24 h; then, (A) lipid peroxidation was measured by TBARS assay, & (B) 4HNE expression was assessed by Western blotting (representative figure, n = 3). In the same cells (C), the protein expression of NRF2 (at 68 kDa) & iNOS were determined by Western blotting (n = 3), while (D) the mRNA expression of catalase (cat) was determined by RT-qPCR (n = 3). (E) The expression of the indicated proteins (pACC, ACC, FOXO1, & PGC-1 beta ) were determined by Western blotting (n = 3, except for PGC-1 beta, where n = 2). (F) 100,000 cells/well 4T1 cells were treated with the indicated concentration of IPA for 24 h; then, the proportions of aldehyde dehydrogenase-positive cells were determined in Aldefluor assays using flow cytometry (n = 3). For Western blots, a typical experiment was displayed. Fold data were log2 transformed to achieve normal distribution. Statistical significance was determined using the ANOVA test followed by Dunnett’s post-hoc test, except for panel F, where Student’s t-test was used. * & *** indicate statistically significant difference between control & treated samples at p < 0.05 & p < 0.001, respectively. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32854297), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
iNOS Antibody - BSA Free

Western Blot: iNOS Antibody - BSA Free [NB300-605] -

Western Blot: iNOS Antibody - BSA Free [NB300-605] - NRF2 activation modulated LCA-induced oxidative stress responses in 4T1 breast cancer cells. The 4T1 cells were treated with 0.3 μm LCA & the NRF2 activator RA839 or tBHQ in the concentrations indicated for 48 h. Lipid peroxidation was determined by measuring (A) TBARS (n = 4) & (B,C) 4-HNE levels using western blotting (n = 3). (D) The 4T1 cells were treated with LCA (0.3 μm) and/or GSH & NAC (both at 5 mm) antioxidants for 48 h, then total protein concentration was determined using the sulforhodamine B assay (n = 3). For statistical analysis ANOVA test was used followed by the Dunnett post-hoc test, where all groups were compared to the LCA-treated cohort. Data are plotted as mean ± SEM. ** p < 0.01, LCA vs. LCA/NRF2-activator-treated groups (GSH, reduced glutathione; LCA, lithocholic acid; NAC, N-acetylcysteine; TBARS, thiobarbituric acid reactive substances; 4HNE, 4-hydroxynonenal). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31461945), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
iNOS Antibody - BSA Free

Western Blot: iNOS Antibody - BSA Free [NB300-605] -

Western Blot: iNOS Antibody - BSA Free [NB300-605] - Impaired amount of virus progeny after stimulation of BV-2 cells with IFN-gamma or IFN-gamma & LPS. BV-2 cells were stimulated with LPS (10 µg/ml), IFN-gamma, (10 ng/ml), LPS + IFN-gamma or IL-4 (10 ng/ml) for 24 h in medium with 2% FBS. Polarization of cells was analyzed by Griess assay (a), Western blot (b) & by detection of the percentage of MHCII+ cells by FACS analysis (c). The amount of virus progeny (pfu/ml) was analyzed by standard plaque assay in the presence of stimulating factors & in normal growth medium in cells & supernatants (d). Statistical analysis was performed related to infection medium. To analyze the recovery of viral replication, normal growth medium was applied after infection of pre-stimulated cells. Standard plaque assay was performed to determine viral progeny in cells & supernatant after 24, 48, 72 & 96 hpi (e). To analyze the relative survival of the cells in the presence of stimulating factors a MTT assay was performed (f). The cellular density of stimulated cells relative to unstimulated (w/o) cells was detected via optical density measurement after 24 & 48 h of cultivation (g). Two-sided t test with unequal variances was used for statistics *p < 0.05, **p < 0.01, ***p < 0.001. Active replication was defined as virus titer above the virus titer of the infection medium (red line). All experiments were performed in triplicate & repeated in an independent experiment. Image collected & cropped by CiteAb from the following publication (http://www.translational-medicine.com/content/13/1/216), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
iNOS Antibody - BSA Free

Western Blot: iNOS Antibody - BSA Free [NB300-605] -

Western Blot: iNOS Antibody - BSA Free [NB300-605] - Inhibitory effect of MMPP on amyloidogenesis & STAT3 translocation in astrocytes & microglia cells. The expression of APP, BACE1 & C99 was detected by Western blotting using specific antibodies in astrocytes (a) & microglia cells (b). Each blot is representative of three experiments. The activity of beta -secretase was investigated using assay kit in astrocytes (c) & microglia cells (d). Values are presented as mean ± S.D. of the three independent experiments performed in triplicate. #p < 0.05 compared to control, *p < 0.05 compared to LPS. Iba-1, COX-2, & iNOS proteins were detected by Western blotting using specific antibodies in astrocytes (e) & microglia cells (f). NO level was measured in astrocytes (g) & microglia cells (h). Activation of STAT3 was investigated using EMSA in astrocytes (i) microglial cells (j) were determined & the expression of STAT3 & phopho-STAT3 was also detected by Western blotting using specific antibodies (k), (l). For the cropped images, samples were run in the same gels under same experimental conditions & processed in parallel. Each band is representative for three experiments Image collected & cropped by CiteAb from the following publication (http://link.springer.com/10.1007/s12017-017-8469-3), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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 5 reviews rated 4.2 using NB300-605 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
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  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

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

P29477 (Mouse)

Additional iNOS Products

Product Documents for iNOS Antibody - BSA Free

Certificate of Analysis

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

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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.

Citations for iNOS Antibody - BSA Free

Customer Reviews for iNOS Antibody - BSA Free (5)

4.2 out of 5
5 Customer Ratings
5 Stars
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20%
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Customer Images

Showing  1 - 5 of 5 reviews Showing All
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  • iNOS protein expression via JESS
    Name: Edma Loku
    Application: Simple Western
    Sample Tested: lung homogenate
    Species: Mouse
    Verified Customer | Posted 09/08/2025
    Lung homogenate protein expression of iNOS in treated and untreated mice
    I tested the protein concentration in my samples using JESS simple western
    iNOS Antibody - BSA Free NB300-605
    Bio-Techne Response
    This review reflects a new species or application tested on a primary antibody.
  • Works okay. it is a bit unspecific
    Name: Edma Loku
    Application: Western Blot
    Sample Tested: mouse lung homogenate
    Species: mouse lung homogenate
    Verified Customer | Posted 09/08/2025
    iNOS protein expression in mouse lung homogenate
    I ran it on my mouse homogenate samples to test the inos response after smoke exposure
    iNOS Antibody - BSA Free NB300-605
  • iNOS Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Liver tissue
    Species: Mouse
    Verified Customer | Posted 09/15/2020
    n/a
    iNOS Antibody - BSA Free NB300-605
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Mouse aortic root
    Species: Mouse
    Verified Customer | Posted 11/13/2015
    Immunofluorescence in mouse paraffin section (RED)
    iNOS Antibody - BSA Free NB300-605
  • Name: Leonid Tarassishin
    Application: Western Blot
    Sample Tested: human astrocyte lysate
    Species: Human
    Verified Customer | Posted 04/28/2014
    iNOS in stimulated astrocytes
    iNOS Antibody - BSA Free NB300-605

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Protocols

View specific protocols for iNOS Antibody - BSA Free (NB300-605):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for iNOS Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

    • Q: We need to evaluate the level of iNOS in Rat sciatic nerve by Western Blot. I see that you have few antibodies suitable for that, could you please recommend the best what you have for our purposes?
      A: We have 4 iNOS antibodies that are suitable for your experiment. The best choice for you will depend on your specific preferences, in this case the main difference will be the immunogen location. If the immunogen is not important to you, then I would recommend choosing NB300-605 as it gives you the best value and we are able to provide the exact immunogen.

    • Q: Would there be any interference from other iNOS isoforms on a Western blot or any other method for NB300-605?
      A: NB300-605 will only recognize the iNOS isoform, so there should be no interference from eNOS, nNOS or bNOS.
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