| Sažetak | Western blot metodom ispitana je ekspresija proteina EXOSC10 u embrionalnim fazama razvoja ribe zebrice (Danio rerio). Protein EXOSC10 sudjeluje u regulaciji ekspresije gena te u procesiranju i degradaciji molekula RNA. Western blot metoda se bazira na specifičnoj interakciji antitijela s proteinom na temelju čega se potvrđuje prisutnost traženog proteina uz određivanje molekulske mase istog. Metoda uključuje redom: pripremu uzoraka, gel elektroforezu, transfer proteina s gela na membranu, inkubiranje primarnim antitijelom, inkubiranje sekundarnim antitijelom i detekciju. Od ranije je poznato da je protein EXOSC10 prisutan u jetri, gonadama, očima i mozgu zebrice, te da je transkript prisutan u prvih 5 dana embrionalnog razvoja. U ovom radu smo pokazali da je protein EXOSC10, molekulske mase oko 100 kDa, prisutan u uzorcima embrija 24 sata, 72 sata i 96 sati nakon fertilizacije. |
| Sažetak (engleski) | The expression of the EXOSC10 protein in the embryonic developmental stages of zebrafish (Danio rerio), whose role is the regulation of gene expression, processes, and degradation of the RNA molecule, was examined using the Western blotting method. The method is based on the specific interaction of the antibody with the protein, on the basis of which the presence of the sought protein is confirmed, and its molecular weight is determined. In detail, the method includes: sample preparation, gel electrophoresis, protein transfer from the gel to a membrane, incubation with primary antibody, incubation with secondary antibody, and detection. It was previously known that the EXOSC10 protein is present in the liver, gonads, eyes, and brain and that the transcript is present in the first 5 days of embryonic development. We have shown that the EXOSC10 protein, with a molecular weight of about 100 kDa, is present in embryo samples at 24 hours, 72 hours, and 96 hours after fertilization. |
