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Uromodulin Canine ELISA

  • Regulatory status:RUO
  • Type:Sandwich ELISA, Biotin-labelled antibody
  • Other names:Tamm-Horsfall urinary glycoprotein, THP, UMOD
  • Species:Canine
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Cat. No. Size Price


RD491163200R 96 wells (1 kit)
PubMed Product Details
Technical Data

Type

Sandwich ELISA, Biotin-labelled antibody

Applications

Serum, Urine

Sample Requirements

5 µl/well

Shipping

At ambient temperature. Upon receipt, store the product at the temperature recommended below.

Storage/Expiration

Store the complete kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box)

Calibration Curve

Calibration Range

0.16–10 ng/ml

Limit of Detection

0.01 ng/ml

Intra-assay (Within-Run)

n = 8; CV = 3.9 %

Inter-assay (Run-to-Run)

n = 6; CV = 7.0 %

Spiking Recovery

Serum: 96.3 %
Urine: 105.3 %

Dilutation Linearity

Serum: 100.3 %
Urine: 102.7 %

Crossreactivity

  • bovine Non-detectable
  • cat Non-detectable
  • dog Non-detectable
  • goat Non-detectable
  • hamster Non-detectable
  • horse Non-detectable
  • human Non-detectable
  • chicken Non-detectable
  • monkey Non-detectable
  • mouse Non-detectable
  • pig Non-detectable
  • rabbit Non-detectable
  • rat Non-detectable
  • sheep Non-detectable
Summary

Features

  • It is intended for research use only
  • The total assay time is less than 3.5 hours
  • The kit measures canine uromodulin in serum and urine
  • Assay format is 96 wells
  • Standard is purified canine native protein (from urine)
  • Components of the kit are provided ready to use, concentrated or lyophilized

Research topic

Renal disease, Animal studies

Summary

Uromodulin, also known as Tamm–Horsfall protein (THP) is the most abundant protein found in mammalian urine under physiological conditions selectively expressed by epithelial cells of the thick ascending limb of Henle’s loop (TALH). It is a glycosylphosphatidylinositol (GPI)-anchored apical membrane protein, released into the tubular lumen through proteolytic cleavage [3]. The normal urinary THP has a molecular weight of 80–90 kDa. Approximately 30% of the molecular weight of THP is carbohydrates, mainly consisting of N-linked complextype
glycans with the most varied array of di-, tri- and tetra-antennary sugar structures. Depending upon the species, urine uromodulin consists of around 640 amino acids. It contains a very high number of cysteine residues that are completely engaged in disulfide bond formation, which is important for the conformation of the protein.
Canine uromodulin is most closely related to human and bovine THP with 80.7 and 79.9% identity to bovine and human THP, respectively. In accordance to humans, the THP in the urine of dogs exists in a polymeric form with molecular weight above 5000 kDa which dissociates into monomeric molecules of around 100 kDa.
It has been suggested that THP protein may be an important component of epithelia, which absorb sodium and chloride ions but are impermeable to water. Uromodulin interacts with other molecules and cells including IL-1, IL-2, TNF, IgG, neuthrophils, lymphocytes and monocytes. THP acts as a host defense factor against urinary tract infections, specifically targeting type 1 fimbriated Escherichia coli. Uromodulin also acts as an inhibitor of stone formation in healthy individuals by trapping crystals in the same manner. However, this function may be subverted under some circumstances and THP may facilitate crystal aggregation and then promote stone formation. It means that it has been proposed to play a dual role as promoter or inhibitor of nephrolithiasis.
A THP-like protein has been studied in dogs specifically because of the species’ urinary excretion of vitamin A. In contrast to humans, carnivores excrete vitamin A in urine as lipophilic retinol and retinyl esters. Therefore, THP is released in the urine of dogs to facilitate the excretion of retinol and retinyl esters.
Previous reports have described urinary THP as biomarker for distal tubular dysfunction in dogs. Its potential use is illustrated by decreased urinary THP in dogs with chronic kidney diseases (CKD) and in dogs with urolithiasis. The high metabolic activity of the cells of the thick ascending limb of Henle’s loop makes them particularly vulnerable to ischemic injury. Consequently, in acute kidney injury (AKI) caused by ischemic insults, evaluation
of THP as an early AKI biomarker is potentially valuable.
The study on the Bernese mountain dogs concluded that THP together with the other identified urinary proteins may be valuable markers for the diagnosis of juvenile nephropathy.

References to Summary

References to Uromodulin

  • Bleyer AJ, Zivna M, Kmoch S. Uromodulin-associated kidney disease. Nephron Clin Pract. 2011;118 (1):c31-6
  • Cavallone D, Malagolini N, Monti A, Wu XR, Serafini-Cessi F. 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
  • Devuyst O, Dahan K, Pirson Y. Tamm-Horsfall protein or uromodulin: new ideas about an old molecule. Nephrol Dial Transplant. 2005 Jul;20 (7):1290-4
  • Gudbjartsson DF, Holm H, Indridason OS, Thorleifsson G, Edvardsson V, Sulem P, de Vegt F, d'Ancona FC, den Heijer M, Wetzels JF, Franzson L, Rafnar T, Kristjansson K, Bjornsdottir US, Eyjolfsson GI, Kiemeney LA, Kong A, Palsson R, Thorsteinsdottir U, Stefansson K. 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
  • Kreft B, Jabs WJ, Laskay T, Klinger M, Solbach W, Kumar S, van Zandbergen G. Polarized expression of Tamm-Horsfall protein by renal tubular epithelial cells activates human granulocytes. Infect Immun. 2002 May;70 (5):2650-6
  • Lau WH, Leong WS, Ismail Z, Gam LH. 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
  • Lhotta K. Uromodulin and chronic kidney disease. Kidney Blood Press Res. 2010;33 (5):393-8
  • Pourmand G, Nasseh H, Sarrafnejad A, Mehrsai A, Hamidi Alamdari D, Nourijelyani K, Shekarpour L. 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
  • Raffi HS, Bates JM Jr, Laszik Z, Kumar S. Tamm-horsfall protein protects against urinary tract infection by proteus mirabilis. J Urol. 2009 May;181 (5):2332-8
  • 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
  • Zaucke F, Boehnlein JM, Steffens S, Polishchuk RS, Rampoldi L, Fischer A, Pasch A, Boehm CW, Baasner A, Attanasio M, Hoppe B, Hopfer H, Beck BB, Sayer JA, Hildebrandt F, Wolf MT. 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
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