Urea Assay Kit ab83362 is a no-wash assay with one 60 min incubation. In the assay, urea is acted on by urease to produce ammonia, followed by the action of glutamate dehydrogenase on ammonia. Readout on any colorimetric (570 nm) microplate reader.
- Complete kit format with full urea assay protocol, and standard curve for quantitation
- Cited in over 100 publications
- Batch sizes up to 500 kits
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- Science advances 10:eadn26892024An ultrathin membrane mediates tissue-specific morphogenesis and barrier function in a human kidney chip.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXingrui Mou,Jessica Shah,Yasmin Roye,Carolyn Du,Samira MusahPubMed 38838141
- Journal of tissue engineering 14:204173142312198132023Rat liver extracellular matrix and perfusion bioreactor culture promote human amnion epithelial cell differentiation towards hepatocyte-like cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSara Campinoti,Bruna Almeida,Negin Goudarzi,Stefan Bencina,Fabio Grundland Freile,Claire McQuitty,Dipa Natarajan,I Jane Cox,Adrien Le Guennec,Vamakshi Khati,Giulia Gaudenzi,Roberto Gramignoli,Luca UrbaniPubMed 38143931
- Nature communications 14:54612023Optogenetic engineering of STING signaling allows remote immunomodulation to enhance cancer immunotherapy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYaling Dou,Rui Chen,Siyao Liu,Yi-Tsang Lee,Ji Jing,Xiaoxuan Liu,Yuepeng Ke,Rui Wang,Yubin Zhou,Yun HuangPubMed 37673917
- PloS one 18:e02894762023The effects of a comparatively higher dose of 1000 mg/kg/d of oral L- or D-arginine on the L-arginine metabolic pathways in male Sprague-Dawley rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDain Raina Kim,Sarah Martin,Kaushik DesaiPubMed 37527267
- Cell proliferation 56:e134652023In vitro construction of liver organoids with biomimetic lobule structure by a multicellular 3D bioprinting strategy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHonglei Jian,Xin Li,Qianqian Dong,Shaonan Tian,Shuo BaiPubMed 37199010
- Toxics 11:2023Modulatory Role of Autophagy in Metformin Therapeutic Activity toward Doxorubicin-Induced Nephrotoxicity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSamar A Antar,Marwa Abd-Elsalam,Walied Abdo,Ahmed Abdeen,Mohamed Abdo,Liana Fericean,Nahed A Raslan,Samah F Ibrahim,Asmaa F Sharif,Amira Elalfy,Hend E Nasr,Ahmed B Zaid,Rania Atia,Ahmed M Atwa,Mohammed A Gebba,Amany A AlzokakyPubMed 36977038
- Current issues in molecular biology 45:1183-11962023Myrrh Essential Oil Mitigates Renal Ischemia/Reperfusion-Induced Injury.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNancy S YounisPubMed 36826023
- Nature communications 13:74972022Targeting endogenous kidney regeneration using anti-IL11 therapy in acute and chronic models of kidney disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAnissa A Widjaja,Sivakumar Viswanathan,Shamini G Shekeran,Eleonora Adami,Wei-Wen Lim,Sonia Chothani,Jessie Tan,Joyce Wei Ting Goh,Hui Mei Chen,Sze Yun Lim,Carine M Boustany-Kari,Julie Hawkins,Enrico Petretto,Norbert Hübner,Sebastian Schafer,Thomas M Coffman,Stuart A CookPubMed 36470928
- Cell reports 41:1117442022Inhibition of glutaminolysis restores mitochondrial function in senescent stem cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDebanik Choudhury,Na Rong,Izuagie Ikhapoh,Nika Rajabian,Georgios Tseropoulos,Yulun Wu,Pihu Mehrotra,Ramkumar Thiyagarajan,Aref Shahini,Kenneth L Seldeen,Bruce R Troen,Pedro Lei,Stelios T AndreadisPubMed 36450260
- Renal failure 44:1819-18322022Salmonella pathogenicity island 1 knockdown confers protection against myocardial fibrosis and inflammation in uremic cardiomyopathy via down-regulation of S100 Calcium Binding Protein A8/A9 transcription.Applications:Unspecified applicationReactive species:Unspecified reactive speciesXinyong Cai,Lang Hong,Yuanyuan Liu,Xiao Huang,Hengli Lai,Liang ShaoPubMed 36299239
Key facts
- Detection method
- Colorimetric
- Sample types
- Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Cell Lysate
- Assay type
- Quantitative
- Range
- 0.5 - 5 nmol/well
- Assay time
- 1h
- Sensitivity
- > 0.5 nmol/well
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Urea Assay Kit ab83362 is a no-wash assay with one 60 min incubation. In the assay, urea is acted on by urease to produce ammonia, followed by the action of glutamate dehydrogenase on ammonia. Readout on any colorimetric (570 nm) microplate reader.
- Complete kit format with full urea assay protocol, and standard curve for quantitation
- Cited in over 100 publications
- Batch sizes up to 500 kits
Key facts
- Detection method
- Colorimetric
- Sample types
- Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Cell Lysate
- Assay type
- Quantitative
- Range
- 0.5 - 5 nmol/well
- Assay time
- 1h
- Assay Platform
- Microplate reader
- Sensitivity
- > 0.5 nmol/well
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
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 Urea Assay Kit II 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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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14 product images
Functional Studies - Urea Assay Kit (ab83362)
Urea measured in mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).
Howie, R Nicole et al., PloS one: vol. 10,7 e0132520., Fig 6, doi:10.1371/journal.pone.0132520Functional Studies - Urea Assay Kit (ab83362)
ab83362 was used as a BUN assay (Blood urea nitrogen assay) to measure urea levels in 80 female Sprague-Dawley rats (age 10-12 months-old).
Functional Studies - Urea Assay Kit (ab83362)
Urea measured in cell culture supernatants and control medium (MEM), background signal subtracted (duplicates +/- SD).
The samples were tested neat; 1:4; 1:16 and 1:64. Only the latter two gave reliable results.
Functional Studies - Urea Assay Kit (ab83362)
Urea measured in cell lysates, background signal subtracted (duplicates; +/- SD).
Functional Studies - Urea Assay Kit (ab83362)
Urea measured in biological fluids, background signal subtracted (duplicates; +/- SD).
Functional Studies - Urea Assay Kit (ab83362)
Example of Urea standard curve using ab83362
Biochemical assay - Urea Assay Kit (ab83362)
Lee et al. used Urea Assay kit ab83362, AST Activity Assay Kit Aspartate Aminotransferase Activity Assay Kit ab105135, and ALT Assay Kit Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 to investigate the pathology in various tissues of K18-hACE2 mice after SARS-CoV-2 infection.
Eight-week-old male K18-hACE2 mice (five mice per group) were infected intranasally with 2 × 103 PFU of SARS-CoV-2 WT (blue symbols) or β variant (B.1.351) (red symbols). Body weight was monitored. Mice were sacrificed at 4 and 6 dpi. Serum and tissue samples were collected. Uninfected mice (−) (black symbols) were used as controls. Multiple-organ damage evaluated by measuring serum markers in WT- and β variant-infected mice at 6 dpi: Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Blood urea nitrogen (BUN). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. NS, not significant. (One-way ANOVA with Tukey’s multiple-comparison test was performed.)
Multiple-organ damage was evaluated by determining the levels of serum AST and ALT (markers of liver injury) and BUN (a marker of kidney injury). AST, ALT, and BUN levels were measured using an AST activity assay kit (no. Aspartate Aminotransferase Activity Assay Kit ab105135), ALT assay kit (no. Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134), and urea assay kit (no. ab83362) (Abcam), respectively, according to the manufacturer’s instructions.
Biochemical assay - Urea Assay Kit (ab83362)
Urea Assay Kit ab83362 used to measure blood urea nitrogen with mouse serum, and Mouse Albumin ELISA kit Mouse Albumin ELISA Kit ab108792 and Creatinine Assay Kit Creatinine Assay Kit ab65340 used to measure albumin-creatinine ratio with mouse urine.
Widjaja et al subjected IL11 null mice and wild type controls to acute kidney injury and ran a blood urea nitrogen assay (Urea Assay Kit ab83362), and measured urine albumin-creatinine ratio using Mouse Albumin ELISA kit Mouse Albumin ELISA Kit ab108792 and Creatinine Assay Kit Creatinine Assay Kit ab65340.
Red indicates acute kidney injury. AKI was induced by intraperitoneal (IP) injection of folic acid (FA, 200 mg/kg) in vehicle (0.3 M NaHCO3) to 10–12-week-old male mice; control mice were administered vehicle alone.
All ELISA and colorimetric assays were performed according to the manufacturer’s protocol.
Biochemical assay - Urea Assay Kit (ab83362)
Antar et al. used Urea Assay kit ab83362 to investigate the effect of metformin therapeutic activity on doxorubicin-induced nephrotoxicity .
Bar-dot plot of the prophylactic effect of Met on DOX-induced renal dysfunction. Prophylactic treatment with Met for 3 weeks decreased urea in the DOX-nephrotoxic mice. Student’s t-test was used for statistical comparisons. Data are expressed as mean ± SEM. Colored dots indicate each individual value and colored curves represent the distribution of each data point. *** p < 0.001 vs. control group and ### p < 0.001 vs. DOX group.
According to the commercially available kits, urea was assessed in accordance with the manufacturer’s instructions (ab83362; Abcam, Cambridge, UK).
Biochemical assay - Urea Assay Kit (ab83362)
Jian et al. used Urea Assay kit ab83362 and Albumin ELISA Kit Human Albumin ELISA Kit ab179887 to investigate liver function markers in a HepaRG/HUVECs/LX‐2‐laden invitro liver model.
Hepatic function evaluations of 2D and 3D models. Determination of the total ALB and urea secretions in the supernatant at days 1, 4 and 7 during culture. The asterisks in figure indicate the level of significant difference (*p < 0.05, **p < 0.01).
For hepatic function measurements, the ALB secretion and urea synthesis were quantified using the corresponding assay kits following the protocol from the manufacturer. At days 1, 4, and 7 during culture, the supernatant was collected and stored at −80°C. The insoluble matter of the sample was centrifugally removed prior to testing. ALB and urea secretions in the supernatant were determined using the ELISA kit (Human Albumin ELISA Kit ab179887, Abcam) and urea assay kit (ab83362, Abcam), respectively. ALB and urea contents were assayed by absorbance at 450 nm and 462 nm on a microplate reader, respectively.
Biochemical assay - Urea Assay Kit (ab83362)
Urea initiates a cascade of reactions ultimately reacting with OxiRed, resulting in a colorimetric product with absorbance at 570 nm.
Biochemical assay - Urea Assay Kit (ab83362)
Diagram showing the principles of the Urea assay method.
Biochemical assay - Urea Assay Kit (ab83362)
Aguilar et al. used Urea Assay kit ab83362 to investigate how the carcinogenic bacterium Helicobacter pylori attaches to gastric cells in a gastric organoid cell culture model.
Quantification of urea concentration in gastric organoid-derived monolayer conditioned media. Fold over control differentiation media (grey, ENRWFGTi_) Organoids were cultured in three different types of media: Orange ENRWFGTiNi. Gray ENRWFGTi_. Green ENR_FGTi_.
The urea concentration in the conditioned-organoid media was measured using a Urea Assay kit (ab83362, Abcam) and following the manufacturer’s instructions. Samples were collected after 24 h, centrifuged for 15 min at 1500 x g to remove cell debris, and diluted 1/100 in the urea kit buffer prior to quantification.
Biochemical assay - Urea Assay Kit (ab83362)
Ramachandra et al. used Urea Assay kit ab83362 to measure urea in brain tissue and to investigate the potential of embelin and levodopa combination therapy for Parkinson’s disease treatment.
Antioxidant potential of embelin (E) and embelin plus LD therapies in mice treated with rotenone, based on the measurement of brain (a) LPO, denoted by MDA, (b) NO, (c) peroxynitrite (PN), and (d) brain urea (BU) levels. The values represent the mean ± standard error (n = 6). For the embelin groups, the number indicates the dose (in mg kg 1 body weight).
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