Methods of protein separation from biological samples. Planning and cloning of an antigen of interest. Production of mono- and polyclonal antisera. Antisera characterization and antibody labelling.
Preparation of samples: collection, fixation and embeding of different biological samples.
Antigen retrieval technics and signal enhancement.
Bases of in situ hibridization and FISH methods.
1. Immunohistochemistry Localization of different cytoskeletal proteins (cytokeratin, tubulin and actin) in the cells of fibroblast culture by using two-steps immunohistochemistry. Fluorescein microscopy and digital microphotography. Localization of a cuticular protein on paraplast-embedded sections from Manduca larvae by using enzyme -labelled second antibody. Light microscopical investigation and microphotography.
2. Electronmicroscopical immunocytochemistry Subcellular localization of heat-shock proteins in normal and pathological samples by using of colloidal gold labelled second antibody. Electronmicroscopical investigation and -photogaphy.
Immunochemical Staining Methods Handbook. Ed. By Th. Boenish. 3rd edition, DAKO, 2001.
David Wild: The Immunoassay Handbook, Newnes, 2013, ISBN 9780080970387