Characterisation of glycans from uromodulin using salt precipitation method

Abdul Rahman Azhari, (2006) Characterisation of glycans from uromodulin using salt precipitation method. Universiti Malaysia Sabah. (Unpublished)

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Abstract

Uromodulin glycoprotein was isolated from urine of pregnant women in different month of pregnancy which are in 9th months and 5th months. The isolation process was done using salt precipitation method originated by Tamm and Horsfall in 1950. When confirming the purity of uromodulin on SDS-page, uromodulin has a molecular weight of 85 to l00 kilo Dalton. The uromodulin isolated from both pregnant women in different months of pregnancy was pure because there are no minor bands formed on the SDS-page gel. Uromodulin glycoprotein was subjected to centrifugation, dialysis and freeze drying to get a dry sample. This project is to characterize the glycans from uromodulin, thus several step was done to examine uromodulin and its glycans. After isolation, steps like reduction and carboxymethylation, tryptic digestion, PNGase F digestion, C�₈ Sep Pak chromatography, reductive elimination and Dowex chromatography was carried out. In addition, experiments like Bradford assay, thin layer chromatography (TLC) separation and serum interaction was also done to examine and observe the characteristic of uromodulin. The result of this experiments showed that the content of uromodulin is high on woman in late pregnancy which is in 9th months and around 9.6 to 10.4 mg/ml after using absorbance of 595 nm in Bradford assay, uromodulin also contain sugar after run on thin layer chromatography plate and uromodulin do not have receptor to interact with serum and therefore not forming agglutination.

Item Type: Academic Exercise
Uncontrolled Keywords: Uromodulin glycoprotein, urine, pregnant, salt precipitation method, isolation, serum interaction
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: SCHOOL > School of Science and Technology
Depositing User: MDM SITI AZIZAH IDRIS
Date Deposited: 05 Feb 2015 03:49
Last Modified: 24 Oct 2017 03:51
URI: http://eprints.ums.edu.my/id/eprint/10396

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