Characterization of glycans rom tamm-horsfall glycoprotein using salt precipitation method

Ahmad Firdaus Haramayni (2006) Characterization of glycans rom tamm-horsfall glycoprotein using salt precipitation method. School of Science Technology. (Unpublished)


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Tamm-Horsfall glycoprotein (THP) is the most abundant protein exists in normal human urine. This glycoprotein can be isolated from the normal human urine by using the Tamm-Horsfall isolation method, which includes the salt-precipitation procedure. This mehod was applied because THP glycoprotein tends to aggregate with salt. The purity of THP glycoprotein was checked by using SDS-PAGE and the size of the protein can be compared with protein marker. From the SDS-PAGE analysis in the presence of reducing agent, the size of THP glycoprotein was in the range of 100 kDa to 120 kDa. The differences between concentration of THP glycoprotein isolated from male and female urine can be figured out by Bradford assay analysis. THP glycoprotein isolated from female urine is higher than THP glycoprotein isolated from male urine. In order to characterize the THP glycoprotein, several methods were used. The characterization of this protein can be achieved by releasing its N-glycans and O-glycans. The release of Nglycans and O-glycans by can be done by treating the THP glycoprotein with trypsin, and further the process with PNGaseF digestion. TLC (Thin Layer Chromatography) method used to determine the carbohydrate contained in both glycans. By doing this method, it showed that galactose existed in N-glycans. Besides, the interaction of THP glycoprotein with serum was also examined. THP glycoprotein showed no agglutination when treated with serum. The serums used in this research were mouse serum, Bovine Serum Albumin (BSA) and Foetal Bovine Serum (FBS).

Item Type: Academic Exercise
Uncontrolled Keywords: glycoprotein, human urine, Tamm-Horsfall isolation method, protein, Thin Layer Chromatography, serum
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: SCHOOL > School of Science and Technology
Date Deposited: 09 Jan 2014 06:45
Last Modified: 10 Oct 2017 07:51

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