Dept. of Immunogenetics

Research programme

The main objective of the department is the study of protein structure and function of cell membranes, particularly molecules which are important in reproductive processes of farm animals. An important tool for identification and analysis of cell surface molecules are monoclonal antibodies. Using the hybridoma technology, we have prepared a set of monoclonal antibodies that are routinely used in our department, and other research institutes in the world for the study of immune functions, including primary and secondary bovine immunodeficiency. Our department aims to identify and characterize CD molecules Experiment_Image214 webprekryv(cluster of differentiation), which act as adhesion molecules, receptors for viruses, signalling molecules or otherwise participating in immune and other processes in the organism. In recent years, the research program of the department has been focused on the study of the function of CD molecules in the reproductive process of cattle. Up to date, although the basic mechanisms of fertilization have been described, all the participating molecules as well as the interactions and the manner of their cooperation in the fertilization process are not yet known. Our research includes a detailed biochemical, histochemical and functional analysis of CD molecules (CD9, CD46 and CD52, and others) on bovine tissues, sperm, oocyte and embryos, changes in their distribution during the development of the reproductive cells as well as explanation of their role in different steps of reproduction. The acquired knowledge can help to understand the immune and reproductive processes in farm animals on the cellular and protein level and allow inter-species comparative studies.

Methodology and Equipment

  • Isolation and cultivation of bovine gametes –capacitation and acrosome reaction of sperm, oocyte maturation, in vitro fertilization- fully equipped and functional sterile box, which includes a laminar box, CO2 incubator, stereomicroscopesoocyt
  • Isolation of blood cells of cattle
  • Immunocytochemical detection of proteins in isolated cells (immunoperoxidase assay, immunofluorescence staining, ELISA)
  • Immunohistochemical analyzes of proteins on frozen tissue sections (Leica cryoctut Cryocut 1800)
  • Biochemical analysis of proteins
  • Electrophoresis- Mini-PROTEAN Tetra Cell (Bio-Rad)
  • Western blot- Trans-Blot® SD Semi-Dry Electrophoretic Transfer Cell (BIO-RAD), and Thermo Scientific Owl™ VEP-2 Mini Tank Electroblotting (Thermo Scientific)
  • 2D electrophoresis (isoelectric focusing) – Multiphor II Electrophoresis system (GE Healthcare)
  • Quick Immunodetection SNAP i.d.® 2.0 Protein Detection System (Millipore)
  • RT-PCR

 

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