Uromodulin--a Useful Marker in Chronic Kidney Disease

Cloud-Clone Corp. Publish date: 2016-06-12

Uromodulin is a glycoprotein encoded by the UMOD gene, which is the most abundant protein in urine of normal mammals. It’s mainly from the Heinz loop of the mammalian kidney.

The molecular weight of monomer UMOD is about 85kDa. UMOD has several different isoforms and exists in aggregates, so the MW could reach millions Da. When the pH is low, precipitations may form.

By the analysis of UMOD sequence, Glu28~Glu149 were chosen for gene cloning and protein expression to get recombinant protein, the cat no. is RPG918Hu01. Immunizing mouse and rabbit with the protein, we got the monoclonal antibody MAG918Hu21, polyclonal antibody PAG918Hu01, biotin labled polyclonal antibody PAG918Hu71. They could be used on WB, ICC, IHC-P, IHC-F, ELISA, etc.

Select high-quality paired antibodies to get the Elisa kit SEG918Hu. It could be used for the detection of UMOD from different species.

Now research shows that there are significant association between UMOD and pregnancy, chronic kidney disease, infertility, etc. The level of UMOD in urine of women with infertility and abortion was significantly higher than that in normal persons. Therefore, urine and sperm are common samples in research. The methods to process urine and semen are listed as follows:

Urine - Aseptically collect the first urine of the day (mid-stream), voided directly into a sterile container. Centrifuge to remove particulate matter, assay immediately or aliquot and store at ≤-20 oC. Avoid repeated freeze-thaw cycles.

Semen-Collect semen samples using a sterile container, then allow it to liquefy at room temperature or 37 oC. After liquefaction, centrifuge at 4000 rpm for 10 min. Remove the supernate and assay immediately or store samples at -20 oC.

The following is the detection data from our lab. The samples detected are urine and serum and the data is shown below.

Table 1: The layout in Elisa

	STD1	STD2	Sample
A	0	0	urine1 100*
B	12.5	12.5	urine2 100*
C	25	25	urine3 100*
D	50	50	urine4 100*
E	100	100	serum1 100*
F	200	200	serum2 100*
G	400	400	serum3 100*
H	800	800	serum4 100*

The dilution fold of samples is 100 fold, and the standards were run in duplicates.

Table 2：The raw data OD in Elisa(450nm-630nm）

	STD1	STD2	Sample
A	0.083	0.091	0.875
B	0.191	0.199	0.245
C	0.237	0.265	1.342
D	0.368	0.348	2.406
E	0.569	0.575	0.545
F	0.943	0.984	0.783
G	1.604	1.738	1.283
H	2.782	2.957	0.562

The samples’ background is not clear, but the data shows good detection performance on samples. And the variances in duplicates is small, and the linearity is over 0.99.

It has been testified for the high quality of the kit. Uromodulin needs further study on clinical function. And the proteins, antibodies and elisa kits developed by Cloud-Clone Corp. offer essential means on uromodulin’s study.

For more information, please enter http://www.cloud-clone.us/.