Ammonia Assay Kit (ab83360)
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
Overview
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Product name
Ammonia Assay Kit
See all Ammonia kits -
Detection method
Colorimetric -
Sample type
Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media -
Assay type
Quantitative -
Sensitivity
20 µM -
Assay time
1h 00m -
Product overview
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 -
Notes
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 medium8References: 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
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Platform
Microplate reader
Properties
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Storage instructions
Store 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 -
Research areas
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Relevance
Ammonia 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.
Images
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Ammonia Assay Kit ab83360 used with cell culture media Li, Zhiyuan et al., PloS one vol. 11,4 e0153526., Fig 1
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.
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Ammonia Assay Kit ab83360 used with liver tissue Wilson, Kyle D et al., Journal of neuroinflammation vol. 15,1 173., Additional file 2Kyle 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.
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Ammonia Assay Kit ab83360 used with cell culture mediaAmmonia 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).
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Ammonia Assay Kit ab83360 used with tissue lysatesAmmonia assay performed with mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).
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Ammonia Assay Kit ab83360 used with lysates of cultured cellsAmmonia assay performed in cell lysates, background signal subtracted (duplicates +/- SD).
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Ammonia Assay Kit ab83360 used with biofluid samplesAmmonia assay performed in biological fluids (range of dilution 1:1-1:100), background signal subtracted (duplicates +/- SD).
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Ammonia assay standard curveExample of ammonia assay standard curve using ab83360
