Ammonia colorimetric assay kit

Manufactured by Abcam

Sourced in United States

The Ammonia Colorimetric Assay Kit is a quantitative biochemical assay used to measure the concentration of ammonia in a variety of sample types. The kit utilizes a colorimetric detection method to quantify the amount of ammonia present in the sample.

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Lab products found in correlation

Ammonia kit buffer, by Abcam (2 mentions) Microplate reader, by Thermo Fisher Scientific (2 mentions) Colorimetric assay kit, by Abcam (1 mentions) Tissuelyser lt, by Qiagen (1 mentions) Bradford reagent, by Bio-Rad (1 mentions)

Spelling variants of this product

Colorimetric assay kit (281 citations) Colorimetric assay (66 citations) Ammonia Assay Kit (28 citations) Ammonia Colorimetric Assay Kit II (3 citations)

11 protocols using ammonia colorimetric assay kit

Plasma and Tissue Ammonia Quantification

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Mouse plasma (25µL) was deproteinized with an equal volume of 8% perchloric acid and centrifuged at 4000g (4°C) for 5 min. Specimens were neutralized with 2M potassium bicarbonate and re-centrifuged at 4000g (4°C) for 10 min prior to analysis. Following the last collection, specimens were analyzed as mentioned above.
Brain and liver tissues were homogenized in 20 times w/v of ice-cold ammonia kit buffer (BioVision^®) and centrifuged at 4000g (4°C) for 5 min. The ammonia concentration was determined using a commercial ammonia colorimetric assay kit (Biovision^®).

Abulseoud O.A., Zuccoli M.L., Zhang L., Barnes A., Huestis M.A, & Lin D.T. (2017). The acute effect of cannabis on plasma, liver and brain ammonia dynamics, a translational study. European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology, 27(7), 679-690.

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Measuring Ammonia Secretion in MCEC Cells

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A total of 1 × 10⁵ MCEC cells/well were plated into 12-well plates in antibiotic-free complete medium overnight, transfected with 50 nM of the indicated siRNAs for 48 hours, and then incubated with an assay media for 20 hours at 33°C. After 68 hours posttransfection, medium supernatant was collected and centrifuged at 500 × g for 5 minutes to remove debris. The supernatant was measured for ammonia concentration with an Ammonia Colorimetric Assay Kit (#K370-100; Biovision, Milpitas, CA, USA).

Choi M, & Bonanno J.A. (2021). Mitochondrial Targeting of the Ammonia-Sensitive Uncoupler SLC4A11 by the Chaperone-Mediated Carrier Pathway in Corneal Endothelium. Investigative Ophthalmology & Visual Science, 62(12), 4.

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Ammonia Quantification in NR-Treated Cells

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Cells (1 x 10⁶ cells per well) were seeded in 24-well plates. For NR treatment, cells were treated with (0.1 mM) NR for 48 h. Ammonia levels were measured by the Ammonia Colorimetric Assay Kit (BioVision, Inc.) according to the manufacturer’s instructions.

Yau W.W., Chen G.B., Zhou J., Francisco J.C., Thimmukonda N.K., Li S., Singh B.K, & Yen P.M. (2023). Nicotinamide riboside rescues dysregulated glycolysis and fatty acid β-oxidation in a human hepatic cell model of citrin deficiency. Human Molecular Genetics, 32(11), 1922-1931.

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Ammonia Quantification in Brain Tissue

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We measured ammonia in the NTS or RVLM tissue lysates by an ammonia colorimetric assay kit (BioVision, Milpitas, CA, USA) according to the manufacturer’s protocol. After the colorimetric reaction, the optical density was read at 570 nm in a micro plate reader (Thermo, Vantaa, Finland).

Tsai C.Y., Su C.H., Chan J.Y, & Chan S.H. (2017). Nitrosative Stress-Induced Disruption of Baroreflex Neural Circuits in a Rat Model of Hepatic Encephalopathy: A DTI Study. Scientific Reports, 7, 40111.

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Measuring Asparaginase Activity in Plasma

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Within 60 min after collection, the blood samples were centrifuged at 4 °C. Samples of plasma were used immediately to measure ammonia (Ammonia Colorimetric Assay Kit, BioVision; Milpitas, CA, USA), following supplier instructions.
Asparaginase activity was measured in plasma samples collected 48 h after infusing the 3rd, 5th, 6th, and 11th doses of Erwinase and for the series of plasma samples collected after infusion of the 12th dose. Activity was based on the rate of ammonia production after adding aliquots of plasma to 37 °C mammalian Ringers with 5 mmol asparagine. The rate of ammonia production was compared to a standard prepared with 0 to 50 U/mL of Erwinase. The standards were linear over this range (r = 0.99 or greater). Asparaginase activity was expressed as U/mL of plasma.

Buddington R.K., Buddington K.K, & Howard S.C. (2022). Multiple Asparaginase Infusions Cause Increasingly Severe Acute Hyperammonemia. Medical Sciences, 10(3), 43.

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Time-Series Plasma Ammonia Assay

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Nine samples were collected in each dosing session; at 0, 2, 4, 6, 8, 10, 15, 30, and 90 min post dose. All samples were collected in pre-cooled heparinized tubes kept in ice water (4°C) during blood collection and centrifuged immediately [3000g (4°C) X 3 min]. Plasma was collected and maintained at 4°C. Immediately following the last collection, specimens were assayed in duplicate using a commercial ammonia colorimetric assay kit (BioVision Inc., Milpitas, CA) according to the manufacturer’s instructions. Ammonia concentration was determined at 570nm wavelength with a Sunrise™ microplate absorbance reader (Tecan Group Ltd., Männerdorf, Switzerland). Inter-assay coefficient of variance of in-house quality controls was <10%.

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Ammonia Quantification in Serum

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Serum ammonia was detected with an Ammonia Colorimetric Assay Kit (Biovision, K370-100) as per the manual. The samples and ammonium chloride standard were added to 96-well plate, bringing the volume to 50 μl/well with assay buffer. Then, 50 μl of the reaction mix was added to each well. The reaction was incubated for 60 min at 37°C and protected from light. The OD 570 nm was measured in a micro plate reader. NH₄Cl concentrations in the samples were calculated according to the NH₄Cl standard curve.

Wu J., Liu J., Lapenta K., Desrouleaux R., Li M.D, & Yang X. (2022). Regulation of the urea cycle by CPS1 O-GlcNAcylation in response to dietary restriction and aging. Journal of Molecular Cell Biology, 14(3), mjac016.

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Plasma and Tissue Ammonia Analysis

Cited 1 time

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We followed the same methods detailed in our previous report. Briefly, rat plasma (25 µL) was deproteinized with an equal volume of 8% perchloric acid and centrifuged at 4000× g (4 °C) for 5 min. Specimens were neutralized with 2 M potassium bicarbonate and re-centrifuged at 4000× g (4 °C) for 10 min prior to analysis. Following the last collection, specimens were analyzed as mentioned above.
Brain and liver tissues were homogenized in 20 times w/v of ice-cold ammonia kit buffer (BioVision^® Milpitas, CA, USA) and centrifuged at 4000× g (4 °C) for 5 min. The ammonia concentration was determined using a commercial ammonia colorimetric assay kit (Biovision^®) [26 (link)].

Badawy A.A., Elghaba R., Soliman M., Hussein A.M., AlSadrah S.A., Awadalla A, & Abulseoud O.A. (2020). Chronic Valproic Acid Administration Increases Plasma, Liver, and Brain Ammonia Concentration and Suppresses Glutamine Synthetase Activity. Brain Sciences, 10(10), 759.

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Hepatic Metabolism Biomarker Assays

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Serum or plasma ammonia levels were measured by an ammonia colorimetric assay kit (BioVision Incorporated; Cat# K370–100) or (Abcam; Cat# ab83360), respectively. Serum or plasma glutamine concentrations were measured by a colorimetric assay kit (BioVision Incorporated; Cat# K556–100). Serum urea content was determined by an assay kit based on Jung’s method (BioVision Incorporated; Cat# K376–100). For blood urea nitrogen (BUN), 10 μl of plasma per sample were pipetted into cups and placed into a Vet Excel Clinical Chemistry Analyzer (Alfa Wassermann Diagnostic Technologies, West Caldwell, NJ). Calculation of urea nitrogen concentration was performed automatically by the clinical chemistry system. Hepatic ATP determination was performed by a fluorometric assay kit according to the manufacturer’s instructions (BioVision Incorporated; Cat# K354–100). Liver lysates were made by homogenization in the corresponding hydrolysis buffer using a TissueLyser LT (Qiagen). Hepatic ATP levels were normalized for protein concentrations determined by Bradford Reagent (Bio-Rad).

Soria L.R., Nitzahn M., Angelis A.D., Khoja S., Attanasio S., Annunziata P., Palmer D.J., Ng P., Lipshutz G.S, & Brunetti-Pierri N. (2019). Hepatic glutamine synthetase augmentation enhances ammonia detoxification.. Journal of inherited metabolic disease, 42(6), 1128-1135.

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Ammonia Metabolism in Embryoid Bodies

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After addition of ammonium chloride, we evaluated ammonia metabolism through changes in ammonia concentration in the cell culture supernatant over a 24-hour period. The standard 1 mM of NH₄Cl was added to the culture dishes containing 100 differentiated embryoid bodies in suspension. We then collected supernatant, and using a colorimetric ammonia assay kit (BioVision, Milpitas, CA), ammonium concentration was measured at 1-, 6- and 24-hour intervals after NH₄Cl addition.

Pettinato G., Lehoux S., Ramanathan R., Salem M.M., He L.X., Muse O., Flaumenhaft R., Thompson M.T., Rouse E.A., Cummings R.D., Wen X, & Fisher R.A. (2019). Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells. Scientific Reports, 9, 8920.

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