Ardi Budi Prakoso (2015) Karakterisasi Protein Cacing Dewasa Eurytrema pancreaticum Dengan Teknik Western_Blot. Skripsi thesis, UNIVERSITAS AIRLANGGA.

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Abstract

The purpose of this research to get the specific protein from adult stage of Eurytrema pancreaticum used Whole Worm Extract (WWE) as antigen which can be used for eurytremiasis diagnostic serologically. E. pancreaticum was collected from ducts pancreas cattle. Amount two hundred adult worms were incubated in incubator with PBS while 12 hours. To homogenize WWE, mortar and stamfer are used, then added by 8 ml of PBS, next WWE homogenized by centrifuge 5000 rpm while 15 minutes. Initiation and booster were done to get policlonal antibody from mice. Identification of protein profile by discontinuous system of SDS- PAGE technique with Coomasie Brilliant Blue staining (12% separating gel and 5% stacking gel). Characterization of spesific protein by semi dry of Western blot technique with BCIP-NBT staining used sera anti-E. pancreaticum and IgG anti- mouse. The result of this research are the protein’s bands which each of them have molecular weight. Identification protein from WWE E. pancreaticum have 24 bands with each of molecular weight of 199, 159, 138, 114, 95, 91, 85, 79, 70, 65, 56, 53, 48, 44, 41, 39, 36, 34, 30, 26, 23, 21, 18 and 14 kDa. The result of characterization of spesific protein from WWE E. pancreaticum have 11 bands with each of molecular weight of 199, 138, 95, 70, 53, 39, 36, 30, 23, 18 and 14 kDa. Band with molecular weight of 53 kDa can be used as antigenic diagnostic for eurytremiasis.

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