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Immunodetection of enzymes catalyzing carbonyl reductions (Western blotting)
Černá, Libuše ; Boušová, Iva (referee) ; Netopilová, Miloslava (advisor)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Libuše Černá Supervisor: PharmDr. Miloslava Netopilová, Ph.D. Title of diploma thesis: Immunodetection of enzymes catalyzing karbonyl reductions (Western blotting) The enzymes which reduce carbonyl group are important for metabolism of eobiotics and xenobiotics. There are recognized three superfamilies of carbonyl- reducing enzymes: aldo-keto reductases (AKR), the medium-chain dehydrogenases/reductases (MDR) and the short-chain dehydrogenases/reductases (SDR). The 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) is member of the SDR superfamily. This enzyme is essential for activating steroid hormone cortison to its active form cortisol. Its activity is also vital for metabolism of some other xenobiotics, e.g the anticancer drug oracin. Yet the carbonyl group of oracin is also reduced by another unknown enzyme wich is located in microsomal membrane. For the purification of this unknown enzyme Q Sepharose was used. The aim was to find out which fractions after purification contain 11β-HSD1 by using the imudetection (Western blotting). In Western blotting we used nitrocelulose membrane, the primary rabbit polyclonal antibodies against 11β-HSD1 dilution 1:1000, the secondary polyclonal...
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Hyaluronan oligosacharides and their biological function
Kučerová, Barbora ; Netopilová, Miloslava (referee) ; Lázníčková, Alice (advisor)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biophysic and Physical Chemistry Candidate: Bc. Barbora Kučerová Supervisor: Doc. Ing. Alice Lázníčková, CSc. Title of diploma thesis: Hyaluronan oligosaccharides and their biological function In my diploma thesis I compile informations about hyaluronan oligosaccharides. Although hyaluronan is in the body found mostly in a form of long chains, we can find it also as small fragments - the oligosaccharides. The oligosaccharides embody different properties than hyaluronan polymers. Hyaluronan oligosaccharides can be synthetised, separated and analysed in laborarory in a variety of ways. Large molecules can be broken into fragments chemically, physically or enzymatically. For thein subsequent separation and purification especially chromatographic methods are used. As the most suitable methods for their final analysis electrophoretic techniques or mass spectrometry were shown. Continous metabolism of hyaluronan proceeds inside the body . Hyaluronan synthase takes a part in synthesis, and the biodegradation important for human is supplyed by several hyaluronidases. Interaction of hyaluronan with cells is mediated by several receptors. Specific receptor for hyaluronan is CD44, which can bind either large polymers and...
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