Griess reagent kit

Manufactured by Dojindo Laboratories

Sourced in Japan

The Griess Reagent Kit is a laboratory product designed to detect and quantify nitrite (NO2-) in various samples. The kit contains the necessary reagents to perform the Griess reaction, which is a colorimetric assay used to measure nitrite levels. The core function of this kit is to provide the necessary tools for researchers and scientists to analyze nitrite concentrations in their samples.

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Lab products found in correlation

Varioskan flash, by Thermo Fisher Scientific (1 mentions) Acetylcholine assay kit, by Cell Biolabs (1 mentions) Enzyme linked immunosorbent assay kit, by R&D Systems (1 mentions) No2 no3 assay kit c 2, by Dojindo Laboratories (1 mentions) Amicon ultra 0.5 ml filter, by Merck Group (1 mentions) Infinite m plex microplate reader, by Tecan (1 mentions) Amicon ultra 0, by Merck Group (1 mentions) Dye binding assay, by Bio-Rad (1 mentions) Infinite m plex, by Tecan (1 mentions)

8 protocols using griess reagent kit

Measuring Nitrite-Producing Activity in Oral Microbiome

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The reaction mixture used to assess NO₂⁻-producing activity contained 100 μL of the dental plaque or tongue coating suspension, 10 µL of 1 M PPB (pH 7.0), 10 μL of 25 mM KNO₃, and 130 μL of de-ionized water. After being subjected to aerobic incubation at 37 °C for 30 min, the reaction mixture was centrifuged at 10,000 rpm at 4 °C for 3 min, and the supernatants were used for the NO₂⁻ assay^{32 (link)}.
The NO₂⁻ concentration was measured with the Griess reagent kit (Dojindo Molecular Technologies, Kumamoto, Japan), in which naphthylethylenediamine dihydrochloride and sulphanilamide reacts with NO₂⁻ to form a purple azo product. The concentration of the azo product was measured colorimetrically at 540 nm using a microplate reader (Varioskan Flash, Thermo Fisher Scientific, Tokyo, Japan).

Sato-Suzuki Y., Washio J., Wicaksono D.P., Sato T., Fukumoto S, & Takahashi N. (2020). Nitrite-producing oral microbiome in adults and children. Scientific Reports, 10, 16652.

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Plasma Biomarkers in Mouse Model

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All mice were euthanized using pentobarbital anesthesia on days zero, seven, 14, and 21. Immediately following euthanization, a 1 ml blood sample was collected from each mouse by cardiac puncture. We measured the plasma acetylcholine concentrations using an Acetylcholine Assay Kit (Cell Biolabs, San Diego, CA, USA). We measured plasma tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) using the respective enzyme-linked immunosorbent assay kits (R&D Systems, Minneapolis, MN, USA and eBioscience Inc., San Diego, CA, USA, respectively) according to the manufacturer's instructions. Plasma NO₂⁻/NO₃⁻ levels were also measured as a surrogate marker for inducible nitric oxide synthase (iNOS) expression using the NO2/NO3 Assay Kit-C II (Colorimetric) and Griess Reagent Kit (DOJINDO Molecular Technologies, Inc., Kumamoto, Japan) for each nitric oxide (NO) radical.

Yokoyama S., Hiramoto K., Yamate Y, & Ooi K. (2017). Influence of Repeated Senna Laxative Use on Skin Barrier Function in Mice. Annals of Dermatology, 29(4), 414-421.

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Nitric Oxide Production in MG6 cells

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MG6 cells cultured in a 96-well plate (3 × 10⁴ cells/well) were pre-treated with hBD1, 2, 3, and 4 at 100 nM and 1 μM for 2 h and then exposed to Pg LPS (10 μg/mL) for 24 h. The supernatant was transferred to a new 96-well plate with Griess reagent (Griess Reagent Kit; Dojindo, Kumamoto, Japan) and incubated at room temperature for 15 min. The amount of nitrite, the major NO metabolite, in the cytosol was measured spectrophotometrically using a microplate reader at a wavelength of 540 nm.

Inoue E., Minatozaki S., Katsuta Y., Nonaka S, & Nakanishi H. (2022). Human β-Defensin 3 Inhibits Porphyromonas Gingivalis Lipopolysaccharide-Induced Oxidative and Inflammatory Responses of Microglia by Suppression of Cathepsins B and L. International Journal of Molecular Sciences, 23(23), 15099.

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Nitric Oxide Quantification in Fungal Cells

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Fungal cells were grown in GMM(Pro) medium, and the stress-treated cell extracts were obtained using the same method described above. After quantifying the amounts of the proteins, the proteins were removed from the cell extracts using Amicon Ultra 0.5-mL filters (Merck, Darmstadt, Germany). The resultant samples were transferred to a 96-well plate, and the NO level was quantified using an NO₂/NO₃ Assay Kit-C II(Colorimetric) ~ Griess Reagent Kit ~ (Dojindo) at 540 nm.

Oiki S., Nasuno R., Urayama S.I., Takagi H, & Hagiwara D. (2022). Intracellular production of reactive oxygen species and a DAF-FM-related compound in Aspergillus fumigatus in response to antifungal agent exposure. Scientific Reports, 12, 13516.

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Arginine-Mediated Nitric Oxide Production

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RAW 264.7 macrophage cell line was seeded into 24-well plate (50 000 cells per well) and incubated for 24 h. After that, the medium was removed, and cells were incubated with fresh Arg deficient medium with or without the pre-stimulation of LPS (1 μg mL⁻¹) for 6 h. Next, cells were incubated with various concentrations of Arg-containing LP for another 48 h. The supernatant of the cultured medium was collected and centrifugated at 1500×g for 15 min. Nitrite (NO₂⁻), long thought to be a biologically inert product of nitric oxide (NO) oxidation, is recognized as a physiological NO storage pool. The amount of NO which reflected by nitrite was quantified by using the Griess Reagent Kit (Dojindo, Kumamoto, Japan) according to the manufacturer's instructions. The absorbance was measured at 540 nm using Infinite M Plex microplate reader (TECAN, Switzerland).

Feng H., Kang J.H., Qi S., Kishimura A., Mori T, & Katayama Y. (2021). Preparation of a PEGylated liposome that co-encapsulates l-arginine and doxorubicin to achieve a synergistic anticancer effect. RSC Advances, 11(54), 34101-34106.

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Measuring Urinary Nitric Oxide Excretion

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Measurement of combined urinary nitrite and nitrate (NOx) excretion is widely used as a marker of nitric oxide (NO) production. Urine samples were collected for 24 h with metabolic cages and were deproteinized by centrifugation with an Amicon Ultra-0.5 filter (EMD Millipore, Darmstadt, Germany). We colorimetrically measured urinary NOx excretion applying the Griess reaction (Griess Reagent Kit; Dojindo, Kumamoto, Japan).^{10 (link)}

Okuyama Y., Hirawa N., Fujita M., Fujiwara A., Ehara Y., Yatsu K., Sumida K., Kagimoto M., Katsumata M., Kobayashi Y., Saka S., Umemura S, & Tamura K. (2017). The effects of anti-hypertensive drugs and the mechanism of hypertension in vascular smooth muscle cell-specific ATP2B1 knockout mice. Hypertension Research, 41(2), 80-87.

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Measurement of Biomarkers in Allergy and Inflammation

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Total protein was measured by dye-binding assay (Bio-Rad Laboratories, Hercules, CA). Allergen specific IgE in the blood of patients was detected by radioallergosorbent testing. IgE, IL-5, IL-4, IL-10, monocyte chemoattractant protein (MCP)-3 were measured using commercial immunoassay kits from BD Biosciences Pharmigen (San Diego, CA). IgG2c was measured using a commercial immunoassay kit from ThermoFisher Scientific (Vienna, Austria). MMP-2, MMP-9, tissue inhibitor of metalloproteinases-2 (TIMP2), IL-13, periostin, and eotaxin were measured using commercial enzyme immunoassay kits from R&D Systems (Minneapolis, MN). The kit used for measuring the plasma levela of MMP-2 in mice measure the active and pro-form of both mouse and human MMP-2 proteins. Nitrite ion in cell supernatant was measured using a Griess reagent kit purchased from Dojindo (Tokyo, Japan). Zymography of MMPs was performed as described (11 (link)).

Takahashi Y., Kobayashi T., D'Alessandro-Gabazza C.N., Toda M., Fujiwara K., Okano T., Fujimoto H., Asayama K., Takeshita A., Yasuma T., Nishihama K., Inoue R., Qin L., Takei Y., Taguchi O, & Gabazza E.C. (2019). Protective Role of Matrix Metalloproteinase-2 in Allergic Bronchial Asthma. Frontiers in Immunology, 10, 1795.

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Nitric Oxide Production in Macrophages

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Macrophages RAW 264.7 or cancer cells were seeded into a 24-well plate (50000 cells/well) and incubated for 24 h. Then, the medium was removed and cells were incubated with a fresh L-arginine deficient medium with or without the stimulation of LPS (1 μg/mL) for 6 h. After that, cells were incubated with various concentrations of Arg for another 48 h. The supernatant of the cultured medium was collected and centrifugated at 1500g for 15 min. The amount of NO, which reflected by nitrite, was quantified by using the Griess Reagent Kit (Dojindo, Kumamoto, Japan) according to the manufacturer's instructions. The absorbance was measured at 540 nm using an Infinite M Plex microplate reader (TECAN, Switzerland).

Feng H., Kishimura A., Mori T, & Katayama Y. (2020). Evaluation of a Synergistic Effect of L-Arginine on the Anticancer Activity of Doxorubicin by Using a Co-culture System. Analytical sciences : the international journal of the Japan Society for Analytical Chemistry, 36(10).

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