Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture media, Serum, Other biological fluids, Cell Lysate,
Assay type:Quantitative,
Sensitivity:= 20 µM,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
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 (λ
= 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 room temperature
- analyze with microplate reader
Technical Note: A dilution series should be performed with urine samples to ensure elimination of any potential matrix effects.
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 lysates
; A549 cell culture medium
- Mouse: brain, liver tissue and serum
; plasma
; pancreas, kidney, liver, brain, and muscle tissue
; faeces
- Bacteria:
B. pertussis
cultures
- Cell culture medium
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
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Ammonia a small molecule with a molecular mass of about 17 daltons is an important player in nitrogen metabolism. It exists primarily as ammonium (NH4+) in physiological pH environments. Ammonia is found predominantly in liver cells where it is produced through amino acid deamination and is further involved in urea production. Ammonia detection can be performed using various tools such as the ammonia detection kit ammonia test kit and the ammonia measurement reader which are essential for researchers to analyze and quantify its presence.
Biological function summary
Ammonia participates in the nitrogen cycle being a substrate for urea synthesis in the liver a vital part of nitrogen waste removal. It combines with carbon dioxide to form carbamoyl phosphate which enters the urea cycle a complex enzymatic system within mitochondria and cytosol. Ammonia's role is not isolated; it interacts with enzymes such as glutamine synthetase. The ammonium assay and ammonia assay provide insights into ammonia’s role and concentration helping elucidate its biological functions and impact.
Pathways
Ammonia is important in the urea cycle and gluconeogenesis. In the urea cycle it is converted to urea preventing toxic accumulation. Gluconeogenesis involves ammonia as it aids in converting pyruvate to glucose a fundamental process for energy maintenance. Related proteins in these pathways include carbamoyl phosphate synthetase in the urea cycle and pyruvate carboxylase in gluconeogenesis. A colorimetric assay might be used to evaluate ammonia's integration in these pathways.
Ammonia plays a significant role in hepatic encephalopathy and hyperammonemia both linked with improper ammonia metabolism. Excess ammonia in the blood due to liver dysfunction can lead to hepatic encephalopathy affecting brain function. Glutamine synthetase activity correlates to ammonia levels impacting the severity of these conditions. Ammonia assay kits are fundamental tools in assessing the association of ammonia with these disorders facilitating better understanding and management of the conditions.
