Testování použitelnosti nově vyvinuté anti-c-Myb protilátky

• Main Author: Pleschnerová, Markéta
• Other Authors: Šnajdr, Pavel
• Format: Dissertation
• Language: Czech
• Published: 2007
• Online Access: http://www.nusl.cz/ntk/nusl-289689

Specific molecules present in cell enables to determine its properties such as function, origin, stage of differentiation. Antigenic features of these molecules are used for their detection, especially in biomedical research and diagnostics. Specific bond between antigen and antibody can be proved by imunological methods e.g. ELISA or Western Blot. Applicability of the antibody for cell and tissue antigens needs to be tested directly by immunocytochemistry and immunohistochemistry. Each antigen requires specific ways of fixation, processing or retrieval. These are the crucial steps for successful antigen detection. Other important steps are appropriate antibody dilution and duration and temperature of incubation. The aim of this work was to determine the applicability of newly developed antic- Myb antibody, obtained by immunization of chick with a fragment of murine c-Myb protein. Blood serum obtained from the same animal before immunization was used as a negative control. Western Blot was applied to verify the specificity of our antibody. Its application for immunocytochemistry was succesfully proved on c-Myb transfected murine embryonic fibroblasts. Results of in situ hybridization (ISH) of c-myb mRNA in whole mount murine embryos compared to published data were used to define prospective c- Myb protein...