Griess assay

Manufactured by Beyotime

Sourced in China

The Griess assay is a colorimetric method used to detect and quantify nitrite (NO2-) levels in various biological samples. It is a widely used analytical technique in research and clinical settings.

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Lab products found in correlation

Daf fm da, by Merck Group (1 mentions) Lsm 710 confocal laser scanning microscope, by Zeiss (1 mentions) Ir780, by Merck Group (1 mentions) Singlet oxygen sensor green (sosg), by Thermo Fisher Scientific (1 mentions) Mitotracker green, by Dojindo Laboratories (1 mentions) Cytotoxicity ros id hypoxia oxidative stress detection kit, by Enzo Life Sciences (1 mentions) Atp assay kit, by Dojindo Laboratories (1 mentions) Mitotracker red, by Beyotime (1 mentions) Daf fm da, by Beyotime (1 mentions) H2dcfda, by Beyotime (1 mentions)

Close spelling variants of this product

Griess assay kit (14 citations) Griess reagent nitric oxide assay kit (2 citations)

9 protocols using griess assay

Quantification of Intracellular ROS and NO

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The intracellular ROS levels were examined using DCFH‐DA (Life Technologies‐Thermo Fisher Scientific) before visualization with a LSM710 Laser Scanning Confocal Microscope (Carl Zeiss) to quantify the fluorescence signals of the oxidized product (2′,7′‐dichlorofluorescein, DCF).
The Griess assay (Beyotime) was used to evaluate the amount of NO in the culture supernatant by measuring the concentration of nitrite (a stable NO breakdown product). An NO⁻ sensitive fluorescence probe DAF‐FM DA (Sigma) was used to detect intracellular NO.22 DAF‐FM DA (10 μmol/L) was used to label the cells at 37°C for 30 minutes before they were washed thrice with PBS. Fluorescence was detected using a LSM710 Laser Scanning Confocal Microscope (scale bars, 100 μm) (Carl Zeiss).

Wang S., Liu F., Tan K.S., Ser H., Tan L.T., Lee L, & Tan W. (2019). Effect of (R)‐salbutamol on the switch of phenotype and metabolic pattern in LPS‐induced macrophage cells. Journal of Cellular and Molecular Medicine, 24(1), 722-736.

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Quantifying Nitric Oxide Production

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To measure NO production, 10 6 Caco-2 cells were plated in 6well dishes and incubated for 21 days after reaching confluence. The concentration of nitrate (NO 3 ) plus nitrite (NO 2 ) was measured in culture supernatants after conversion of NO 3 to NO 2 by nitrate reductase, with detection of NO 2 using a commercial kit (Griess assay; Beyotime Institute of Biotechnology, Jiangsu, China).

Zeng J., Jiang J., Zhu W, & Chu Y. (2016). Heat-killed yogurt-containing lactic acid bacteria prevent cytokine-induced barrier disruption in human intestinal Caco-2 cells. Annals of microbiology, 66(1).

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Quantifying ET and NO Secretion

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Cells were seeded at 5×10⁵ cells/well into 6-cm culture dishes and treated for 24 or 48 h before 100 μl of the supernatant was collected from each well and centrifuged to remove impurities and cell debris. ET was detected by ELISA. A standard curve was obtained via serial dilution of standards included in the assay kit (Beyotime Institute of Biotechnology). Each sample was aliquoted into a microtiter plate and OD at 450 nm was measured. NO was detected using a Griess assay (Beyotime Institute of Biotechnology), carried out in accordance with the manufacturer's instructions and with OD measured at 540 nm using a microplate spectrophotometer.

Shangguan W.J., Zhang Y.H., Li Z.C., Tang L.M., Shao J, & Li H. (2017). Naringin inhibits vascular endothelial cell apoptosis via endoplasmic reticulum stress- and mitochondrial-mediated pathways and promotes intraosseous angiogenesis in ovariectomized rats. International Journal of Molecular Medicine, 40(6), 1741-1749.

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Nitric Oxide Secretion in MIN6 Cells

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MIN6 cells were seeded in 96‐well microtiter plates and treated with media, IFN‐γ (100 ng/ml) or LIGHT (5 μg/ml) alone or in combination for 48 h. Nitric oxide (NO) level in culture supernatant was determined by measuring the levels of nitrite, a stable by‐product of NO by using Griess assay (Beyotime Institute of Biotechnology) according to the manufacturer's suggested protocols.

Zheng Q., Cao Z., Hu X., Li G., Dong S., Xu G, & Zhang K. (2016). LIGHT/IFN‐γ triggers β cells apoptosis via NF‐κB/Bcl2‐dependent mitochondrial pathway. Journal of Cellular and Molecular Medicine, 20(10), 1861-1871.

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Measuring Nitric Oxide in Chondrocytes

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The nitric oxide (NO) levels in the culture supernatant were detected using the Griess assay (Beyotime, Shanghai, China; S0021S), following the manufacturer's protocol. The chondrocytes were cultured overnight in 96-well plates (3 × 10⁴/well). After the cells were treated with Q3GA and IL-1β, 50 μl of the cell supernatant was transferred into a new 96-well plate, followed by addition of 50 μl of Griess Reagent I and 50 μl of Griess Reagent II. Absorbance was measured at 540 nm using a microplate reader.

Mao H., Feng Y., Feng J., Yusufu Y., Sun M., Yang L, & Jiang Q. (2024). Quercetin-3-O-β-D-glucuronide attenuates osteoarthritis by inhibiting cartilage extracellular matrix degradation and inflammation. Journal of Orthopaedic Translation, 45, 236-246.

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Nitrite Production Assay in Lung Cells

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Nitrite production was assayed by measurement of the nitrite ion concentration with the Griess assay (Beyotime Institute of Biotechnology, Shanghai, China) according to the manufacturer’s protocol. Lung CD45⁺ cells (1 × 10⁶ cells), cDC1s, CD24⁻CD11b⁺ DC2s, CD24⁺ cDC2s, MCs, AMs and IMs (5 × 10⁴ cells) were sorted from mice 1.5 days after the first OVA challenge and cultured for 48 h. NO production was measured in the supernatants of the cultured cells.

Yi S., Zhai J., Niu R., Zhu G., Wang M., Liu J., Huang H., Wang Y., Jing X., Kang L., Song W., Shi Y, & Tang H. (2018). Eosinophil recruitment is dynamically regulated by interplay among lung dendritic cell subsets after allergen challenge. Nature Communications, 9, 3879.

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Quantifying Microglial Nitric Oxide

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Murine primary microglia were co-cultured with GBM cells GL261 with different treatments for 24 h. Aliquots of cell culture supernatant were taken at 24 h for total NO production by using the Griess assay according to the protocol of the manufacturer (Beyotime Institute Biotechnology, Jiangmen, China), as absorbance at 540 nm from the ELISA plate reader [11 (link)]. The amount of nitrite was normalized to the amount produced by untreated microglia.

Yin Y., Qiu S., Li X., Huang B., Xu Y, & Peng Y. (2017). EZH2 suppression in glioblastoma shifts microglia toward M1 phenotype in tumor microenvironment. Journal of Neuroinflammation, 14, 220.

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Nitric Oxide Production in Macrophages

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Briefly, RAW264.7 cells or primary PMs (1.5 × 10⁴ cells/100 μl) were plated in 96-well plates overnight and treated with various positive control drugs and concentrations of DCL prior to stimulation of LPS (0.5 μg/ml) and IFNγ (10 ng/ml) for 24 h. The NO production in cells were detected immediately by using the Griess assay according to the protocol of the manufacturer (Beyotime, Shanghai, China).

Yuan Y., Hu Q., Liu L., Xie F., Yang L., Li Y., Zhang C., Chen H., Tang J, & Shen X. (2022). Dehydrocostus Lactone Suppresses Dextran Sulfate Sodium-Induced Colitis by Targeting the IKKα/β-NF-κB and Keap1-Nrf2 Signalling Pathways. Frontiers in Pharmacology, 13, 817596.

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Oxidative Stress and Hypoxia Detection

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IR780 was purchased from Sigma Corporation. Griess assay, H₂DCFDA, Mitotracker Red and DAF-FM DA were obtained by Beyotime Institute of Biotechnology (China). Singlet Oxygen Sensor Green (SOSG) was purchased from Thermo Fisher Scientific. Cytotoxicity ROS-ID Hypoxia/Oxidative Stress Detection Kit was obtained from Enzo Life Sciences. The 2-iminothiolane, isopentylnitrite, perfluorodecalin (FDC) and fluorescein isothiocyanate (FITC) were obtained from Aladdin Industrial Corporation (Shanghai, China). Mitotracker Green, Mitochondrial Membrane Potential Detection Kit (JC-1), and ATP assay kit were purchased from Dojindo Laboratories (Japan). All of the chemicals used in this study were of analytical grade and used without further purification.

Ji C., Si J., Xu Y., Zhang W., Yang Y., He X., Xu H., Mou X., Ren H, & Guo H. (2021). Mitochondria-targeted and ultrasound-responsive nanoparticles for oxygen and nitric oxide codelivery to reverse immunosuppression and enhance sonodynamic therapy for immune activation. Theranostics, 11(17), 8587-8604.

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