Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Cell Lysate,
Assay type:Quantitative,
Sensitivity:> 0.5 nmol/well,
Range:0.5 - 5 nmol/well,
Assay time:30m,
Assay Platform:Microplate reader
Product details:
Urea Assay Kit ab83362 is a rapid, simple, sensitive, and reliable assay used to quantify urea in a variety of samples such as serum, plasma, and urine, etc. It can be used as a blood urea nitrogen assay kit (BUN assay kit).
How the assay works
In the urea assay protocol, urea is acted on by enzymes to form a product that reacts with a probe to generate color (ODmax=570nm). The absorbance is directly proportional to the concentration of urea in the solution. The kit can detect as low as 0.5 nmol per well or 10 μM of urea.
Urea assay protocol summary
- Add samples and standards to wells
- Add reaction mix and incubate for 60 min at room temperature
- Analyze with microplate reader
Urea assay methods
There are three urea assay methods to run blood urea nitrogen assays that are commonly used in biological research:
a) Jung's method based on o-phthalaldehyde and N-(1-naphthyl)ethylenediamine, in which o-phthalaldehyde reacts with urea, followed by a reaction with N-(1-naphthyl)ethylenediamine, or an alternative chemical, to form a colored product. Jung's method is simple, with the addition of a single working reagent, with a short incubation at room temperature followed by colorimetric readout. However the reagents include hazardous sulfuric acid.
Abcam provides Urea Assay Kit
ab234052
which is based on an improved version of Jung's method.
b) An enzymatic assay based on the conversion of urea to ammonium and carbon dioxide by urease, followed by measurement of the ammonium level by reaction with Berthelot's reagent (an alkaline solution of phenol and hypochlorite) to form a colored product, or an alternative method of ammonium readout
c) An enzymatic assay based on the conversion of urea to ammonium and carbon dioxide by urease, followed by the conversion of ammonium and NADH to glutamate and NAD+ by glutamate dehydrogenase. The assay can then either readout by measuring the decrease in the 340 nm absorbance of NADH, or by use of further enzymes to produce a colored product that increases proportionate to the amount of urea in the sample.
Urea Assay Kit ab83362 is based on the glutamate dehydrogenase method, with further enzymes, to allow a positive readout.
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K375 Urea Colorimetric Assay Kit. K375-100 is the same size as the 100 test size of ab83362.
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.
Urea also known as carbamide is a small molecule with a molecular mass of 60.06 g/mol. It results from protein metabolism and is eliminated by the kidneys. Urea plays a central role in the nitrogen cycle in humans. The liver predominantly expresses it where it is then transported to the kidneys for excretion. Blood urea nitrogen (BUN) and plasma urea levels are measured using assays like the BUN kit and the BUN test Boston to assess kidney function.
Biological function summary
In the context of nitrogen waste management urea serves as an essential substance in detoxifying ammonia. Urea is part of the urea cycle a series of biochemical reactions converting ammonia into urea. Ammonia results from amino acid breakdown and its accumulation can be toxic. The urea cycle ensures safe excretion therefore playing a protective role in maintaining nitrogen balance.
Pathways
The urea cycle is interlinked with the citric acid cycle especially in the liver. It involves critical enzymes such as carbamoyl phosphate synthetase and ornithine transcarbamylase. These pathways share intermediates and energy regulation roles ensuring efficient functioning of nitrogen metabolism. The pathways connect with proteins like arginase which facilitate the final step in the conversion of arginine to urea.
Improper urea cycle function leads to conditions such as hyperammonemia and acute renal failure. Elevated blood urea nitrogen levels indicate potential kidney dysfunction. Moreover defects in proteins like arginase or ornithine transcarbamylase can disrupt this balance resulting in urea cycle disorders. Monitoring blood urea levels helps in diagnosing and managing these conditions effectively.
