Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Cell culture media, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 20 µM
Product nameAmmonia Assay Kit
See all Ammonia kits
Sample typeUrine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
Assay time1h 00m
Ammonia Assay Kit (ab83360) provides a rapid, simple, sensitive, and reliable assay for ammonia and ammonium.
In the ammonia assay protocol, ammonia and ammonium are converted to a product that reacts with a probe to generate color (λmax = 570 nm) which can be easily quantified by plate reader. The kit can detect 1 nmol (~20 µM) of total ammonia and ammonium, which is much more sensitive than measuring ammonia with a NADPH based assay.
Ammonia assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 60 min at 37ºC
- analyze with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K370 Ammonia Colorimetric Assay Kit. K370-100 is the same size as the 100 test size of ab83360.
Ammonia assays measure the total level of the ammonium ion and of ammonia in a sample.
At a physiological pH, nearly all ammonia exists as ammonium in solution.
The chemical equation that drives the relationship between ammonia and ammonium is: NH3 + H2O ↔ NH4+ + OH-
When the pH is low, the reaction is driven to the right, and when the pH is high, the reaction is driven to the left. In general, at around room temperature, at a pH less than 6.0, the proportion of ammonium-N plus ammonia-N as NH3 is very low and as NH4+ is very high. At a pH around 8.0 (the ammonia assay buffer pH), the proportion as NH3 is 10 percent or less.
This is our most popular ammonia assay kit; ammonia assay kit ab102509 uses an alternative non-enzymatic method.
How other researchers have used Ammonia Assay Kit ab83360
The ammonia assay kit has been used in publications in a variety of sample types, including:
- Human: A549, 293T and U2OS cell culture lysates1; A549 cell culture medium2
- Mouse: brain, liver tissue and serum3; plasma4; pancreas, kidney, liver, brain, and muscle tissue5; faeces6
- Bacteria: B. pertussis cultures7
- Cell culture medium8
References: 1 - Xu S et al 2019, Spinelli JB et al 2017, Trempolec N et al 2017; 2 - Jin L et al 2018; 3 - Wilson et al 2018; 4 - Khoja S et al 2018, Cantero et al 2016; 5 - Gutierrez-de-Jua V et al 2017; 6 - Shen et al 2016; 7 - Fyson et al 2017; 8 - Li et al 2016
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests Ammonia Assay Buffer WM 1 x 25ml Converting Enzyme (Lyophilized) Blue 1 vial Developer Orange 1 vial Enzyme Mix (Lyophilized) Green 1 vial NH4Cl Standard Yellow 1 x 100µl OxiRed Probe in DMSO Red 1 x 200µl
RelevanceAmmonia is an important source of nitrogen for living systems. Nitrogen is required for the synthesis of amino acids, which are the building blocks of protein. Ammonia is a metabolic product which is created through amino acid deamination. It plays an important role in both normal and abnormal animal physiology like normal animal acid/base balance.
Zhiyuan Li et al. used ammonia assay kit ab83360 to examine the build-up of ammonia in old cell culture medium containing glutamine and how this might induce autophagy in cultured cells.
The ammonia assay was used to measure the ammonia concentration in cell culture medium containing Glutamine (medium alone, without cells) which was stored at 4°C or at 37°C for between 1-7 days. They found that ammonia levels increased significantly in the medium stored at 37°C.
Kyle Wilson et al. used ammonia assay kit ab83360 to examine ammonia levels in liver tissue in response to P. chabaudi infection. They found that in IL-10 KO mice, ammonia levels were not elevated above WT during P. chabaudi infection.
WT and IL-10 KO mice were infected with P. chabaudi and monitored during the peak of infection. WT mice were sacrificed at the peak of infection (day 10 p.i.) and IL-10 KO mice upon severe morbidity as determined via SHIRPA score.
Ammonia assay performed with cell culture medium (range of dilution 1:10-1:200) and control medium (range of dilution 1:1-1:20), background signal subtracted (duplicates +/- SD).
Ammonia assay performed with mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).
Ammonia assay performed in cell lysates, background signal subtracted (duplicates +/- SD).
Ammonia assay performed in biological fluids (range of dilution 1:1-1:100), background signal subtracted (duplicates +/- SD).
Example of ammonia assay standard curve using ab83360
ab83360 has been referenced in 23 publications.
- Sivangala Thandi R et al. Ornithine-A urea cycle metabolite enhances autophagy and controls Mycobacterium tuberculosis infection. Nat Commun 11:3535 (2020). PubMed: 32669568
- Lu Z et al. Functional Changes of the Community of Microbes With Ni-Dependent Enzyme Genes Accompany Adaptation of the Ruminal Microbiome to Urea-Supplemented Diets. Front Microbiol 11:596681 (2020). PubMed: 33414773
- Nitzahn M et al. Split AAV-Mediated Gene Therapy Restores Ureagenesis in a Murine Model of Carbamoyl Phosphate Synthetase 1 Deficiency. Mol Ther 28:1717-1730 (2020). PubMed: 32359471
- Xu S et al. An Integrative Systems Biology and Experimental Approach Identifies Convergence of Epithelial Plasticity, Metabolism, and Autophagy to Promote Chemoresistance. J Clin Med 8:N/A (2019). PubMed: 30736412
- Khoja S et al. A constitutive knockout of murine carbamoyl phosphate synthetase 1 results in death with marked hyperglutaminemia and hyperammonemia. J Inherit Metab Dis N/A:N/A (2019). PubMed: 30835861