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Original title:
Biochemical methods as tool for study of reproductive proteins
Authors: Postlerová, Pavla ; Zigo, Michal ; Pohlová, Alžběta ; Jonáková, Věra
Document type: Papers
Conference/Event: XXIst SYMPOSIUM OF BIOLOGY AND IMMUNOLOGY OF REPRODUCTION WITH INTERNATIONAL PARTICIPATION, Třešť (CZ), 2015-05-14 / 2015-05-16
Year: 2015
Language: eng
Abstract: Study of molecular mechanisms in reproduction is essential for the understanding of this outstanding process. Our lab studies proteins secreted by reproductive organs and sperm using various biochemical methods for a long time. We have expertise in protein extraction from spermatic cells using different approaches, and by kits for proteins from the sperm surface and distinct subcellular compartments. The proteins of reproductive organ fluids are separated by chromatographic methods, such as size exclusion chromatography, high-performance liquid chromatography with reverse phase (RP-HPLC) and affinity chromatography on matrices with various ligands. Proteins are subjected to SDS- or 2D-electrophoresis for their characterization and comparison of various extraction methods, different mammalian species, and sperm in different functional development. Electrophoretically separated proteins may be transferred onto nitrocellulose membrane (Western blot) for antibody detection or binding studies with lectin-labelled ligands (lectins, polysaccharides, zona pellucida glycoproteins). We use immunoprecipitation method with specific antibody for protein determination followed by the MALDI identification. Proteins are localized by immunofluorescent techniques on/in spermatic cells and tissue sections of reproductive organs. Isolation of proteins from reproductive tissues and fluids, and the antibody detection is crucial for the studying of reproductive protein origin.
Keywords: HPLC; lectins; monoclonal antibody; zona pellucida glycoproteins
Project no.: GA P503/12/1834, GA 14-05547S, ED1.1.00/02.0109
Funding provider: GA ČR, GA ČR, GA MŠk
Host item entry: Book of abstracts of XXIst Symposium of biology and immunology of reproduction
Institution: Institute of Biotechnology AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0270356
Permalink: http://www.nusl.cz/ntk/nusl-263524
The record appears in these collections:
Research > Institutes ASCR > Institute of Biotechnology
Conference materials > Papers
Authors: Postlerová, Pavla ; Zigo, Michal ; Pohlová, Alžběta ; Jonáková, Věra
Document type: Papers
Conference/Event: XXIst SYMPOSIUM OF BIOLOGY AND IMMUNOLOGY OF REPRODUCTION WITH INTERNATIONAL PARTICIPATION, Třešť (CZ), 2015-05-14 / 2015-05-16
Year: 2015
Language: eng
Abstract: Study of molecular mechanisms in reproduction is essential for the understanding of this outstanding process. Our lab studies proteins secreted by reproductive organs and sperm using various biochemical methods for a long time. We have expertise in protein extraction from spermatic cells using different approaches, and by kits for proteins from the sperm surface and distinct subcellular compartments. The proteins of reproductive organ fluids are separated by chromatographic methods, such as size exclusion chromatography, high-performance liquid chromatography with reverse phase (RP-HPLC) and affinity chromatography on matrices with various ligands. Proteins are subjected to SDS- or 2D-electrophoresis for their characterization and comparison of various extraction methods, different mammalian species, and sperm in different functional development. Electrophoretically separated proteins may be transferred onto nitrocellulose membrane (Western blot) for antibody detection or binding studies with lectin-labelled ligands (lectins, polysaccharides, zona pellucida glycoproteins). We use immunoprecipitation method with specific antibody for protein determination followed by the MALDI identification. Proteins are localized by immunofluorescent techniques on/in spermatic cells and tissue sections of reproductive organs. Isolation of proteins from reproductive tissues and fluids, and the antibody detection is crucial for the studying of reproductive protein origin.
Keywords: HPLC; lectins; monoclonal antibody; zona pellucida glycoproteins
Project no.: GA P503/12/1834, GA 14-05547S, ED1.1.00/02.0109
Funding provider: GA ČR, GA ČR, GA MŠk
Host item entry: Book of abstracts of XXIst Symposium of biology and immunology of reproduction
Institution: Institute of Biotechnology AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0270356
Permalink: http://www.nusl.cz/ntk/nusl-263524
The record appears in these collections:
Research > Institutes ASCR > Institute of Biotechnology
Conference materials > Papers
Record created 2017-03-17, last modified 2021-11-24