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Urea assay kit

Manufactured by Merck Group
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The Urea Assay Kit is a laboratory diagnostic tool used to measure the concentration of urea in biological samples, such as blood, urine, or other fluids. It provides a quantitative analysis of urea levels, which is an important indicator of kidney function and nitrogen metabolism.

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Lab products found in correlation

Bcg albumin assay kit, by Merck Group (2 mentions) Creatinine assay kit, by Merck Group (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Negative sirna sc 37007, by Santa Cruz Biotechnology (1 mentions) E cadherin, by Proteintech (1 mentions) Ripa lysis buffer, by Cell Signaling Technology (1 mentions) Supersignal chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Arginase, by Merck Group (1 mentions) Bicinchoninic acid protein assay kit, by CWBIO (1 mentions) Colorimetric detection kit, by Thermo Fisher Scientific (1 mentions) Mak080, by Merck Group (1 mentions) Activity detection kit, by Solarbio (1 mentions) Glucose oxidase method, by Thermo Fisher Scientific (1 mentions) Porcine insulin radioimmunoassay kit, by Merck Group (1 mentions) Pico tag reverse phase column, by Waters Corporation (1 mentions) Protein assay dye reagent concentrate solution, by Bio-Rad (1 mentions) V plex kit, by Mesoscale (1 mentions) Analytical balance, by Mettler Toledo (1 mentions) Filter paper, by Cytiva (1 mentions) Beaker glass, by Iwaki (1 mentions)

22 protocols using urea assay kit

1

Molecular Mechanisms of Cisplatin-Induced Kidney Injury

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MM102 (HY-12220A) and PFA (HY-15484) were purchased from MedChemExpress (NJ, USA). Cisplatin was purchased from the Pharmacy of Rhode Island Hospital (NDC 0703-5747-11, RI, USA). Antibodies to C-cas3 (9664), MLL1 (14197), p53 (2524), p-p53 (ser15) (9284), ATM (2873), p-ATM (4526), ATR (2790), p-ATR (2853), Chk1 (2360), p-Chk1 (2348), Chk2 (2662), p-Chk2 (2661), and RIPA lysis buffer (9806) were purchased from Cell Signaling Technology (MA, USA). Antibodies to WDR5 (ab56919), histone H3 (ab1791), histone H2AX (ab124781), and γ-H2AX (ab81299) were from Abcam (MA, USA). E-cadherin was bought from Proteintech (20874-1-AP, IL, USA). Antibody to NGAL was purchased from R&D systems (AF1857, MN, USA). MLL1 siRNA (Assay ID 501621), Lipofectamine 2000 (11668019), and SuperSignal chemiluminescent substrate (34580) were from Thermo Fisher Scientific (MA, USA). H3K4me3 antibody (07-473), TUNEL Kit (11684795910), CCK8 (96992), DAPI (D9542), Creatinine Assay Kit (6M01K06250), and Urea Assay Kit (7C08K03750) were purchased from Sigma-Aldrich (MO, USA). Klotho antibody (sc-515942), WDR5 siRNA (sc-61799), E-cadherin siRNA (sc-35243), p53 siRNA (sc-29436), and negative siRNA (sc-37007) were purchased from Santa Cruz Biotechnology (CA, USA).
Zhang C., Guan Y., Zou J., Yang X., Bayliss G, & Zhuang S. (2022). Histone methyltransferase MLL1 drives renal tubular cell apoptosis by p53-dependent repression of E-cadherin during cisplatin-induced acute kidney injury. Cell Death & Disease, 13(9), 770.
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2

Arginase-Coupled Assay for Argininosuccinate Lyase Activity

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ASL activity was detected by a coupled assay using arginase and measuring urea production as described previously [15 (link)]. First, 50 μl of cell extract was mixed with 100 μl of 13.6 mM argininosuccinate in water and 100 μl of arginase (50 units) (Sigma-Aldrich, St. Louis, MO, USA) in 66.7 mM PBS (pH 7.5). The reactions were stopped after 30 min by adding perchloric acid (2% final concentration). Then, a 10-μl aliquot of the stopped reaction was sampled to assay the urea concentration with a urea assay kit (Sigma-Aldrich). The ASL activities of the wild-type and mutant proteins were expressed as the concentration of urea/the total protein concentration in mU/mg protein. The total protein concentration was assayed by using the Bicinchoninic acid protein assay kit (CWBIO, Beijing, China). All experiments were performed in triplicate.
Zhao M., Hou L., Teng H., Li J., Wang J., Zhang K, & Yang L. (2019). Whole-Exome Sequencing Identified a Novel Compound Heterozygous Genotype in ASL in a Chinese Han Patient with Argininosuccinate Lyase Deficiency. BioMed Research International, 2019, 3530198.
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3

Serum Biomarker Quantification in Mice

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Mouse blood samples were collected at the end of experiments. Serum urea, urea nitrogen (BUN), and creatinine were respectively assessed by Urea assay kit (MAK006, Sigma-Aldrich, St. Louis, MO, USA), Urea Nitrogen (BUN) colorimetric detection kit (EIABUN, Thermo Fisher Scientific, Waltham, MA), and Creatinine assay kit (MAK080, Sigma-Aldrich) according to the manufacturer’s instructions.
Hong Y., Wang J., Sun W., Zhang L., Xu X, & Zhang K. (2023). Gallic acid improves the metformin effects on diabetic kidney disease in mice. Renal Failure, 45(1), 2183726.
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4

Microbial Nitrogen Metabolism Analysis

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Ammonia concentration was determined in the supernatant using an automated phenol-hypochlorite method (Broderick and Kang, 1980 (link)). The concentration of urea was determined using a urea assay kit (Sigma-Aldrich) based on the manufacturer’s protocol. The concentration of total free α-amino acids in the supernatant of the mixed amino acid-grown cultures was analyzed using ninhydrin as previously described (Jones et al., 2002 (link)). Microbial crude protein (MCP) was determined by the Folin phenol method (Makkar et al., 1982 (link)). Urease and glutamine synthase (GS) activity were determined using urease and glutamine synthase assay kits (Sigma-Aldrich), respectively, and glutamate dehydrogenase (GDH) and glutamate synthetase (GOGAT) activity were measured using activity detection kits (Solarbio, Beijing, China) according to the manufacturer’s instructions.
Hailemariam S., Zhao S, & Wang J. (2020). Complete Genome Sequencing and Transcriptome Analysis of Nitrogen Metabolism of Succinivibrio dextrinosolvens Strain Z6 Isolated From Dairy Cow Rumen. Frontiers in Microbiology, 11, 1826.
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5

Postprandial Metabolic Biomarkers in Blood

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Blood levels of circulating insulin, glucose, urea and free AA were measured on day 8. Blood was sampled from the jugular vein immediately before feeding, and at 30, 60, 90, 120, 180, and 240 min post-feeding. Samples were centrifuged at 12000×g for 2 min within 10 min of collection, and plasma was stored at −20 °C for subsequent analyses. Plasma insulin concentrations were measured using a porcine insulin radioimmunoassay kit (Millipore, St. Charles, MO, USA). Plasma glucose levels were determined with the glucose oxidase method (Thermo Scientific, Waltham, MA, USA). Determination of free AA concentration in plasma was performed by high-performance liquid chromatography (PICO-TAG reverse-phase column; Waters, Milford, MA, USA) using an analytical method based on deproteinization and derivatization of AA with phenylisothiocyanate (Burrin et al. 1995 (link)). Plasma urea levels were measured at 0 and 120 min using a commercial urea assay kit (Sigma-Aldrich, St Louis, MO, USA).
Manjarín R., Columbus D.A., Suryawan A., Nguyen H.V., Hernandez-García A.D., Hoang N.M., Fiorotto M.L, & Davis T. (2015). Leucine supplementation of a chronically restricted protein and energy diet enhances mTOR pathway activation but not muscle protein synthesis in neonatal pigs. Amino acids, 48(1), 257-267.
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6

Evaluating Hepatic Spheroid Function

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Spheroid albumin and urea levels were examined to assess HepG2 and HepaRG spheroid liver-like functionality (BCG Albumin Assay Kit, MAK124 and Urea Assay Kit, MAK006, Sigma Aldrich, UK). All assays were performed as per manufacturer’s instructions. However, to ensure that samples fell within the standard curve, for the urea assay, the supernatants were diluted 1:10 with urea assay. On the day of harvest, spheroids were pooled and centrifuged at 230g for 5 min. Cell culture supernatant was collected (1.5 ml) for use with the albumin and urea secretion assays. Samples were stored at -−80°C until the assays were performed.
Conway G.E., Shah U.K., Llewellyn S., Cervena T., Evans S.J., Al Ali A.S., Jenkins G.J., Clift M.J, & Doak S.H. (2020). Adaptation of the in vitro micronucleus assay for genotoxicity testing using 3D liver models supporting longer-term exposure durations. Mutagenesis, 35(4), 319-330.
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7

Comprehensive Urinary and Plasma Analysis

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Urinary protein was determined by Bradford method using Protein Assay Dye Reagent Concentrate solution (Bio-Rad). Albumin and creatinine concentrations were measured using corresponding kits that were purchased from Sigma. Blood plasma samples were collected in EDTA-treated tubes by centrifugation at 2,000× g for 20 minutes to deplete platelets, and inflammation-related biomarkers were assayed using the V-PLEX kit (Meso Scale Discovery). In addition, blood urea nitrogen was measured using urea assay kit (Sigma). All procedures were performed according to manufacturer instructions.
Hsu M.F., Bettaieb A., Ito Y., Graham J., Havel P.J, & Haj F.G. (2017). Protein tyrosine phosphatase Shp2 deficiency in podocytes attenuates lipopolysaccharide-induced proteinuria. Scientific Reports, 7, 461.
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8

Passion Fruit Peel Bioactive Extraction

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The materials used in this research were purple passion fruit peel, red passion fruit peel, yellow passion fruit peel, ethanol (E-Merck), ethylene diamine tetraacetic acid (E-Merck), alanine transaminase activity assay kit (Sigma Aldrich), aspartate transaminase activity assay kit (Sigma Aldrich), urea assay kit (Sigma Aldrich), creatinine assay kit (Sigma Aldrich), sodium carboxymethyl cellulose (Tokyo Chemical Industry), and demineralized water (Brataco).
The tools used in this research were cutter (Kenko), drying cabinet (Alumex), blender (Miyako), balance (Mettler Toledo), analytical balance (Mettler Toledo), filter paper (Whatman), dropper (Iwaki), evaporating dish (Iwaki), maceration vessel (Iwaki), beaker glass (Iwaki), measuring glass (Iwaki), volumetric flask (Iwaki), test tube (Iwaki), hot plate stirrer (Thermo), water bath (Memmert), incubator (Memmert), rotary evaporator (Buchi), thermometer (Lutron), test sieve 10 (Retsch), stopwatch (Casio), spectrophotometer (Agilent).
Nerdy N, & Ritarwan K. (2019). Hepatoprotective Activity and Nephroprotective Activity of Peel Extract from Three Varieties of the Passion Fruit (Passiflora Sp.) in the Albino Rat. Open Access Macedonian Journal of Medical Sciences, 7(4), 536-542.
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9

Biochemical Markers Analysis in Metabolic Disorders

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Plasma triglycerides, total cholesterol, and high-density lipoprotein cholesterol (HDL-C) were measured by spectrophotometric methods, as described by Bucolo et al., [44 (link)] and using SpinReact Kits (SpinReact S.A., Girona, Spain). Glucose concentration was evaluated by an Ascensia ELITE XL blood glucose meter (Bayer Consumer Care AG, Basel, Switzerland). Plasma insulin was measured using a Rat/Mouse Insulin ELISA kit (Millipore Corporation, Billerica, MA, USA) following the manufacturer’s instructions. Hemoglobin concentration was measured according to the Drabkin method [45 (link)], and the percentage of hematocrit was calculated in capillary tubes after centrifugation. Plasma urea levels were determined by using the Urea Assay Kit (Sigma, St. Louis, MO, USA) according to manufacturer’s instructions. Plasma non-enzymatic antioxidant capacity was measured as the oxygen radical absorbance capacity (ORAC) [46 (link)]. Oxidized and reduced glutathione balance (GSSG/GSH) was assayed according to Hissin and Hilf [47 (link)].
Méndez L., Muñoz S., Barros L., Miralles-Pérez B., Romeu M., Ramos-Romero S., Torres J.L, & Medina I. (2023). Combined Intake of Fish Oil and D-Fagomine Prevents High-Fat High-Sucrose Diet-Induced Prediabetes by Modulating Lipotoxicity and Protein Carbonylation in the Kidney. Antioxidants, 12(3), 751.
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10

Hepatocyte Culture and Metabolite Analysis

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Human primary hepatocytes were cultured in serum-free hepatocyte defined growth medium as described in section “Human primary hepatocyte culture”. Hepatocyte culture media was collected at indicated time points and stored at -20 °C before analysis. The concentration of urea was measured using urea assay kit and the concentration of albumin was assessed using BCG (Bromocresol Green) albumin assay kit (both Sigma-Aldrich) according to manufacturer’s instructions.
Klaas M., Möll K., Mäemets-Allas K., Loog M., Järvekülg M, & Jaks V. (2021). Long-term maintenance of functional primary human hepatocytes in 3D gelatin matrices produced by solution blow spinning. Scientific Reports, 11, 20165.
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