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Nitric Oxide (NO) Colorimetric Assay Kit

Citations(45)

  • Cat.No.:E-BC-K035-M

  • Detection instrument: Microplate reader(540-550 nm)

Manual MSDS

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  • 48T
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Detection principle

NO is easily oxidized to form NO2- in vivo or in aqueous solution, and a reddish azo compound is formed with the color developing agent, and the concentration of the azo compound is linearly related to the concentration of NO. The concentration of NO can be calculated indirectly by measuring the OD value at 550 nm.

Performance characteristics

Synonyms NO
Sample type Serum, plasma, tissue, saliva
Sensitivity 0.16 μmol/L
Detection range 0.16-100 μmol/L
Detection method Colorimetric method
Assay type Quantitative
Assay time 55 min
Precision Average inter-assay CV: 3.700%Average intra-assay CV: 2.400%
Other instruments required Micropipettor, Vortex mixer, Centrifuge
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
Storage Reagent 6: -20℃, others: 2-8℃
Valid period 12 months

Dilution of sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.16-100 μmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type

Dilution factor

Human serum

1

Human plasma

1

10% Mouse liver tissue homogenization

1

Rat serum

1

Rat plasma

1

10% Epipremnum aureum tissue homogenization

1

Note: The diluent is double distilled water, normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).


Citations

  1. Frontiers in Immunology (2021) IF: 7.561
    Transcriptional Profiling and Functional Analysis of N1/N2 Neutrophils Reveal an Immunomodulatory Effect of S100A9-Blockade on the Pro-Inflammatory N1 Subpopulation

    DOI: 10.3389/fimmu.2021.708770
    Sample: neutrophils
  2. Biomaterials Science (2022) IF: 7.59
    Transdermal delivery of allopurinol on acute hyperuricemic mice via polymer microneedles for regulation of serum uric acid levels

    DOI: 10.1039/D2BM01836E
    Sample: serum
  3. Antioxidants (2022) IF: 7.675
    Therapeutic Effects of Heterotrigona itama (Stingless Bee) Bee Bread in Improving Hepatic Lipid Metabolism through theActivation of the Keap1/Nrf2 Signaling Pathway in an Obese Rat Model

    DOI: 10.3390/antiox11112190
    Sample: liver
  4. SCIENCE OF THE TOTAL ENVIRONMENT (2022) IF: 7.963
    Cold exposure, gut microbiota, and hypertension: A mechanistic study

    DOI: 10.1016/j.scitotenv.2022.155199

    PMID: 35417730

    		 Sample: plasma
  5. FOOD AND CHEMICAL TOXICOLOGY (2021) IF: 6.025
    Reactive astrocytes induced by 2-chloroethanol modulate microglia polarization through IL-1β, TNF-α, and iNOS upregulation

    DOI: 10.1016/j.fct.2021.112550

    PMID: 34517076

    		 Sample: astrocyte
  6. Antioxidants (2021) IF: 6.313
    Hepatoprotective Effect of Bee Bread in Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) Rats: Impact on Oxidative Stress and Inflammation

    DOI: 10.3390/antiox10122031

    PMID: 34943134

    		 Sample: liver
  7. BIOMEDICINE & PHARMACOTHERAPY (2020) IF: 6.53
    Gastroprotective effect of gallic acid against ethanol-induced gastric ulcer in rats: Involvement of the Nrf2/HO-1 signaling and anti-apoptosis role

    DOI: 10.1016/j.biopha.2020.110075

    PMID: 32179202

    		 Sample: Tissue homogenate
  8. BIOMEDICINE & PHARMACOTHERAPY (2021) IF: 6.53
    D-ribose-L-cysteine prevents oxidative stress and cardiometabolic syndrome in high fructose high fat diet fed rats

    DOI: 10.1016/j.biopha.2021.112017

    PMID: 34399203

    		 Sample: heart
  9. Oxidative Medicine and Cellular Longevity (2021) IF: 6.543
    Leonurine Ameliorates Oxidative Stress and Insufficient Angiogenesis by Regulating the PI3K/Akt-eNOS Signaling Pathway in H2O2-Induced HUVECs

    DOI: 10.1155/2021/9919466
    Sample: HUVECs
  10. HYPERTENSION (2018) IF: 6.823
    Potential Protective Mechanism in the Cardiac Microvascular Injury

    DOI: 10.1161/HYPERTENSIONAHA.118.11035
    Sample: cardiac microvascular endothelial cells (CMECs)
  11. INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES (2021) IF: 6.953
    Structural characterization and immunomodulatory activities of two polysaccharides from Rehmanniae Radix Praeparata

    DOI: 10.1016/j.ijbiomac.2021.06.100

    PMID: 34197855

    		 Sample: RAW264.7 cells
  12. Marine Drugs (2021) IF: 5.118
    MMP-9 and IL-1β as Targets for Diatoxanthin and Related Microalgal Pigments: Potential Chemopreventive and Photoprotective Agents

    DOI: 10.3390/md19070354

    PMID: 34206447

    		 Sample: HaCaT cells,B16-F0 cells
  13. Nutrients (2022) IF: 5.719
    Paraxanthine Supplementation Increases Muscle Mass, Strength, and Endurance in Mice

    DOI: 10.3390/nu14040893

    PMID: 35215543

    		 Sample: serum
  14. Frontiers in Pharmacology (2021) IF: 5.811
    Identification of the Antithrombotic Mechanism of Leonurine in Adrenalin Hydrochloride-Induced Thrombosis in Zebrafish via Regulating Oxidative Stress and Coagulation Cascade.

    DOI: 10.3389/fphar.2021.742954
  15. Frontiers in Pharmacology (2022) IF: 5.811
    Effects of Pereskia aculeate Miller Petroleum Ether Extract on Complete Freund’s Adjuvant-Induced Rheumatoid Arthritis in Rats and its Potential Molecular Mechanisms

    DOI: 10.3389/fphar.2022.869810
    Sample: RAW264.7
  16. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 5.924
    Parathyroid Hormone Induces Human Valvular Endothelial Cells Dysfunction That Impacts the Osteogenic Phenotype of Valvular Interstitial Cells

    DOI: 10.3390/ijms23073776

    PMID: 35409134

    		 Sample: valvular endothelial cell
  17. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2021) IF: 5.924
    Bee Bread Ameliorates Vascular Inflammation and Impaired Vasorelaxation in Obesity-Induced Vascular Damage Rat Model: The Role of eNOS/NO/cGMP-Signaling Pathway

    DOI: 10.3390/ijms22084225

    PMID: 33921777

    		 Sample: Tissue homogenate(aortic )
  18. Frontiers in Pharmacology (2022) IF: 5.988
    Anti-arthritic and toxicological evaluation of ethanolic extract of Alternanthera bettzickiana in rats.

    DOI: 10.3389/fphar.2022.1002037
    Sample: serum
  19. ACS Omega (2022) IF: 4.132
    Ajuga bracteosa Exerts Antihypertensive Activity in l-NAME-Induced Hypertension Possibly through Modulation of Oxidative Stress, Proinflammatory Cytokines, and the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway

    DOI: 10.1021/acsomega.2c03888

    PMID: 36157749

    		 Sample: serum
  20. Food Bioscience (2021) IF: 4.24
    Auricularia polytricha and Flammulina velutipes ameliorate inflammation and modulate the gut microbiota via regulation of NF-κB and Keap1/Nrf2 signaling pathways on DSS-induced inflammatory bowel disease

    DOI: 10.1016/j.fbio.2021.101426
    Sample: colon,liver
  21. JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
    Ziziphus Oxyphylla hydro-methanolic extract ameliorates hypertension in L-NAME induced hypertensive rats through NO/cGMP pathway and suppression of oxidative stress related inflammatory biomarkers.

    DOI: 10.1016/j.jep.2021.114825
    Sample: Serum
  22. JOURNAL OF ETHNOPHARMACOLOGY (2022) IF: 4.36
    Aronia melanocarpa (Michx.) Elliott. attenuates dextran sulfate sodium-induced Inflammatory Bowel Disease via regulation of inflammation-related signaling pathways and modulation of the gut microbiota

    DOI: 10.1016/j.jep.2022.115190

    PMID: 35306040

    		 Sample: liver homogenates
  23. Journal of Functional Foods (2021) IF: 4.451
    Structural characterization and immunomodulatory activity of a glucan from Radix Codonopsis

    DOI: 10.1016/j.jff.2021.104537
    Sample: Cell culture supernatant
  24. Andrology (2022) IF: 4.456
    Icariin inhibits the formation of mitochondria-associated membranes (MAMs) and improves erectile function in rats treated with prostate radiation

    DOI: 10.1111/andr.13218

    PMID: 35765205

    		 Sample: penile cavernous tissue
  25. MOLECULES (2023) IF: 4.927
    Neuroprotective Effects of Savinin on LPS-Induced Neuroinflammation In Vivo via Regulating MAPK/NF-κB Pathway and NLRP3 Inflammasome Activation

    DOI: 10.3390/molecules28041575
    Sample: hippocampa
  26. INTERNATIONAL IMMUNOPHARMACOLOGY (2022) IF: 4.932
    Daurisoline alleviated experimental colitis in vivo and in vitro: Involvement of NF-κB and Wnt/β-Catenin pathway

    DOI: 10.1016/j.intimp.2022.108714

    PMID: 35366641

    		 Sample: colon tissues,RAW 264.7 cell
  27. JOURNAL OF ETHNOPHARMACOLOGY (2018) IF: 3.115
    Anti-inflammatory action of Athyrium multidentatum extract suppresses the LPS-induced TLR4 signaling pathway

    DOI: 10.1016/j.jep.2018.02.031
    Sample: peritoneal macrophages (PMs)
  28. Animals (2022) IF: 3.231
    Expression of Hormones, Cytokines, and Antioxidants in Heat-Stressed Subfertile Female Dromedaries

    DOI: 10.3390/ani12162125

    PMID: 36009715

    		 Sample: serum
  29. BEHAVIOURAL BRAIN RESEARCH (2022) IF: 3.352
    Prenatal chronic stress impairs the learning and memory ability via inhibition of the NO/cGMP/PKG pathway in the Hippocampus of offspring

    DOI: 10.1016/j.bbr.2022.114009

    PMID: 35850398

    		 Sample: hippocampal tissue
  30. Andrology (2022) IF: 3.842
    Low androgen status inhibits erectile function by upregulating the expression of proteins of mitochondria-associated membranes in rat corpus cavernosum

    DOI: 10.1111/andr.13188

    PMID: 35466583

    		 Sample: penile cavernous tissue
  31. Andrology (2021) IF: 3.842
    Low androgen status inhibits erectile function by increasing pyroptosis in rat corpus cavernosum

    DOI: 10.1111/andr.12995

    PMID: 33657666

    		 Sample: Cell lysate
  32. Andrology (2022) IF: 3.842
    Low androgen level impairs erectile function of rat by regulating the Ng/CaN/AKT/eNOS pathway in penile corpus cavernosum

    DOI: 10.1111/andr.13202

    PMID: 35678097

    		 Sample: penile cavernous tissue
  33. Veterinary Sciences (2021) IF: 2.304
    Protective and Therapeutic Effects of Orlistat on Metabolic Syndrome and Oxidative Stress in High-Fat Diet-Induced Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) in Rats: Role on Nrf2 Activation

    DOI: 10.3390/vetsci8110274
    Sample: liver
  34. JOURNAL OF MOLECULAR HISTOLOGY (2021) IF: 2.611
    Salivary exosomes as a new therapy to ameliorate diabetes mellitus and combat xerostomia and submandibular salivary glands dysfunction in diabetic rats

    DOI: 10.1007/s10735-020-09935-z

    PMID: 33389429

    		 Sample: serum
  35. DNA AND CELL BIOLOGY (2018) IF: 2.634
    GPR30 Attenuates Myocardial Fibrosis in Diabetic Ovariectomized Female Rats: Role of iNOS Signaling

    DOI: 10.1089/dna.2018.4208

    PMID: 30227089

    		 Sample: Serum
  36. Iranian Journal of Basic Medical Sciences (2021) IF: 2.699
    Corilagin potential in inhibiting oxidative and inflammatory stress in LPS-induced murine macrophage cell lines (RAW 264.7)

    DOI: 10.22038/IJBMS.2021.59348.13174
    Sample: RAW 264.7 cell
  37. ACS Omega (2020) IF: 2.87
    Antiarthritic Potential of Comprehensively Standardized Extract of Alternanthera bettzickiana: In Vitro and In Vivo Studies

    DOI: 10.1021/acsomega.0c01670

    PMID: 32803042

    		 Sample: Serum
  38. Molecular Medicine Reports (2021) IF: 2.952
    α‑rhamnrtin‑3‑α‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway

    DOI: 10.3892/mmr.2021.12439

    PMID: 34523697

    		 Sample: RAW264.7 cell
  39. MOLECULAR BIOLOGY REPORTS (2020) IF: 1.402
    Codeine-induced hepatic injury is via oxido-inflammatory damage and caspase-3-mediated apoptosis

    DOI: 10.1007/s11033-020-05983-6

    PMID: 33211294

    		 Sample: lvier
  40. IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL (2019) IF: 1.645
    Adventitious root cultures of Oplopanax elatus inhibit LPS-induced inflammation via suppressing MAPK and NF-κB signaling pathways

    DOI: 10.1007/s11626-019-00396-7
    Sample: PMs cells
  41. Journal of Obsessive-Compulsive and Related Disorders (2021) IF: 1.677
    TGF-β1, neopterin, tetrahydrobiopterin, and nitric oxide levels in pediatric obsessive–compulsive disorder

    DOI: 10.1016/j.jocrd.2021.100649
    Sample: Serum
  42. BIOMEDICAL AND ENVIRONMENTAL SCIENCES (2019) IF: 1.917
    Effects of Simulated Heat Wave and Ozone on High Fat Diet ApoE Deficient Mice

    DOI: 10.3967/bes2018.101
    Sample: Plasma,Tissue homogenate
  43. ANDROLOGIA (2020) IF: 1.951
    Low androgen status inhibits erectile function by up‐regulating the expression of P2X receptors in rat corpus cavernosum

    DOI: 10.1111/and.13627
    Sample: corpus cavernosum
  44. Journal of Clinical Tuberculosis and Other Mycobacterial Diseases (2021)
    Phagocytosis and the antigen-processing abilities of macrophages derived from monocytes in spinal tuberculosis patients

    DOI: 10.1016/j.jctube.2021.100215
    Sample: macrophages
  45. Advances in Traditional Medicine (2020)
    Anti-hypernociceptive effects of methanol extract of Boswellia dalzielii on STZ-induced diabetic neuropathic pain

    DOI: 10.1007/s13596-019-00411-y
    Sample: serum,sciatic nerves
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Reviews/Q&A

  • Show all
  • Reviews
  • Q&A
Q

H**********dSubmitted [ Mar 13 2023 ]

Asked:  We followed the following procedure, but the kit couldn\'t detect the NO: The amount of nitrite in the culture medium was measured as an indicator of NO production. The amount of nitrite was calculated using NO assay kit reagent (Catalog No: E-BC-K035-M). Then, 100μL of the culture supernatant was added to 100 μL of NO reagent and incubated for 10 minutes in a dark room. The absorbance was measured at 550 nm on a microplate reader. The nitrite concentration was calculated using the sodium nitrite standard curve. Could you please guide us?

Reply

A

adminSubmitted [ Mar 13 2023 ]

Answered: Hi Hafedh Ahmed, Thanks for your inquiry. I have checked your procedure which does not correspond to the steps in our manual. And I sent you the real manual by email. After you checked the manual, pls give us feedback. Thank you

Q

H***************iSubmitted [ Mar 10 2023 ]

Asked: Can the kit use for RAW 264.7 cells?

Reply

A

adminSubmitted [ Mar 10 2023 ]

Answered: Hi, Hafedh. Thanks for your inquiry. For Nitric Oxide (NO) Colorimetric Assay Kit, you can use catalog number E-BC-K135-M to detect cell lines. E-BC-K135-M Nitric Oxide (NO) Colorimetric Assay Kit (Nitrate Reductase)

Q

M*****aSubmitted [ Aug 22 2022 ]

Asked: The reagent 5 is crystallized, does it affect the result after melting?

Reply

A

adminSubmitted [ Aug 22 2022 ]

Answered: Reagent 5 is a kind of pure acid reagent with high concentration, which is easy to crystallize in cold storage. It is OK to balance it to room temperature before use, which can not affect the result.

Q

K*****nSubmitted [ Dec 14 2020 ]

Asked: Can the kit use for HUVEC Cell? How much does it ship to Thailand. Thanks.

Reply

A

adminSubmitted [ Dec 14 2020 ]

Answered: Hi Korawan, Sorry to tel you this kit can't test cell samples. Any questions, pls feel free to ask. Best regards,

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