Uromodulin Human ELISA

Summary

Uromodulin (Tamm-Horsfall protein, UMOD) is approx. 85-kDa glycoprotein that is produced in the thick ascending limb of Henle´s loop and early distal convoluted tubules of the nephron. It is a transmembrane protein, which is secreted into the urine through proteolytic cleavage of the glycosylphospha­tidylinositol (GPI) anchor. It belongs to the GPI family. Healthy individuals excrete tens of miligrams of uromodulin per day, making in the most abundant protein in the urine. Uromodulin modulates cell adhesion and signal transduction by interacting with cytokines and it inhibits the aggregation of calcium crystals. By reducing calcium oxalate precipitation, uromodulin plays a protective role with respect to renal stone formation as demonstrated by recent studies on THP- deficient mice prone to nephrolithiasis. THP acts as a host defense factor against urinary tract infections induced by uropathogens such as Esherichia coli, Staphylococcus saphrophyticus, Proteus mirabilis and Klebsiela pneumonie. Uromodulin binds to type 1 fimbriae of Escherichia coli and thereby blocks colonization of urothelial cells. Tamm-Horsfall protein interacts with other molecules and cells including IL-1, IL-2, TNF, IgG, neuthrophils, lymphocytes and monocytes. Binding of uromodulin to neutrophils induces synthesis of IL-8, provokes the respiratory burst and degranulation and stimulates chemotaxis and phagocytosis. Recently, genome-wide association studies identified uromodulin as a risk factor for chronic kidney disease and hypertension. Mutations in the Uromodulin gene are associated with three autosomal dominant tubulo-interstitial nephropathies such as familial juvenile hyperuricemic nephropathy (FJHN), medullary cystic kidney disease (MCKD2) and glomerulocystic kidney disease (GCKD). These disorders are characterized by juvenile onset of hyperuricemia, gout and progressive renal failure.

Features

• It is intended for research use only
• The total assay time is less than 3.5 hours
• The kit measures uromodulin in serum, plasma (EDTA, citrate, heparin) and urine
• Assay format is 96 wells
• Standard is native protein based
• Quality Controls are human serum based
• Components of the kit are provided ready to use, concentrated or lyophilized

Research topic

Renal disease

Find out more on biovendor.com

• SERUM UROMODULIN A SCREENING MARKER OF NEPHROPATHY in BioVendor News

Assay format

Sandwich ELISA, Biotin-labelled antibody

Applications

Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum, Urine

Sample requirements

10 µl

Storage/Shipping

2–8°C

Calibration Curve

Calibration range

0.5 to 32 ng/ml

Limit of detection

0.12 ng/ml

Intra-assay (Within-Run, n=8)

CV = 2.0 %

Inter-assay (Run-to-Run, n=6)

CV = 6.4 %

Spiking Recovery

95.7 %

Dilution Linearity

99.9 %

Cross-Reactivity

References to summary

• Beck BB, Hoppe B, Attanasio M, Boehm CW, Baasner A, Fischer A, Pasch A, Rampoldi L, Polishchuk RS, Steffens S, Wolf MT, Hildebrandt F, Boehnlein JM, Hopfer H, Zaucke F, Sayer JA. Uromodulin is expressed in renal primary cilia and UMOD mutations result in decreased ciliary uromodulin expression. Hum Mol Genet. 2010 May 15;19 (10):1985-97
• Bleyer AJ, Zivna M, Kmoch S. Uromodulin-associated kidney disease. Nephron Clin Pract. 2011;118 (1):c31-6
• Gam LH, Ismail Z, Leong WS, Lau WH. Qualification and application of an ELISA for the determination of Tamm Horsfall protein (THP) in human urine and its use for screening of kidney stone disease. Int J Biol Sci. 2008;4 (4):215-22
• Holm H, Sulem P, Stefansson K, Gudbjartsson DF, Thorsteinsdottir U, Palsson R, Kiemeney LA, Kong A, Eyjolfsson GI, Bjornsdottir US, Kristjansson K, Rafnar T, Wetzels JF, Franzson L, den Heijer M, d'Ancona FC, de Vegt F, Indridason OS, Edvardsson V, Thorleifsson G. Association of variants at UMOD with chronic kidney disease and kidney stones-role of age and comorbid diseases. PLoS Genet. 2010 Jul;6 (7):e1001039
• Jabs WJ, Kreft B, van Zandbergen G, Kumar S, Solbach W, Laskay T, Klinger M. Polarized expression of Tamm-Horsfall protein by renal tubular epithelial cells activates human granulocytes. Infect Immun. 2002 May;70 (5):2650-6
• Kumar S, Laszik Z, Bates JM Jr, Raffi HS. Tamm-horsfall protein protects against urinary tract infection by proteus mirabilis. J Urol. 2009 May;181 (5):2332-8
• Lhotta K. Uromodulin and chronic kidney disease. Kidney Blood Press Res. 2010;33 (5):393-8
• Monti A, Malagolini N, Cavallone D, Serafini-Cessi F, Wu XR. Variation of high mannose chains of Tamm-Horsfall glycoprotein confers differential binding to type 1-fimbriated Escherichia coli. J Biol Chem. 2004 Jan 2;279 (1):216-22
• Nourijelyani K, Pourmand G, Shekarpour L, Hamidi Alamdari D, Nasseh H, Mehrsai A, Sarrafnejad A. Urinary Tamm-Horsfall protein and citrate: a case-control study of inhibitors and promoters of calcium stone formation. Urol J. 2005 Spring;2 (2):79-85
• Pirson Y, Dahan K, Devuyst O. Tamm-Horsfall protein or uromodulin: new ideas about an old molecule. Nephrol Dial Transplant. 2005 Jul;20 (7):1290-4
• Rhodes DC. Binding of Tamm-Horsfall protein to complement 1q and complement 1, including influence of hydrogen-ion concentration. Immunol Cell Biol. 2002 Dec;80 (6):558-66
• Zasloff M. Antimicrobial peptides, innate immunity, and the normally sterile urinary tract. J Am Soc Nephrol. 2007 Nov;18 (11):2810-6

By research topic:

• Renal disease

By molecule:

• Uromodulin

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