3 articles published in JoVE
A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts Valentin Dunsing1, Salvatore Chiantia1 1Institute for Biochemistry and Biology, Cell Membrane Biophysics Group, University of Potsdam This protocol describes a fluorescence fluctuation spectroscopy-based approach to investigate interactions among proteins mediating cell-cell interactions, i.e. proteins localized in cell junctions, directly in living cells. We provide detailed guidelines on instrument calibration, data acquisition and analysis, including corrections to possible artefact sources.
Generation of Murine Monoclonal Antibodies by Hybridoma Technology Pamela Holzlöhner1, Katja Hanack1 1Department of Biochemistry and Biology, University of Potsdam An optimized protocol is presented for the generation of monoclonal antibodies based on the hybridoma technology. Mice were immunized with an immunoconjugate. Spleen cells were fused by PEG and an electric impulse with immortal myeloma cells. Antibody-producing hybridoma cells were selected by HAT and antigen-specific ELISA screening.
Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis Verónica A. Lombardo*1,2,3, Cécile Otten*1,2, Salim Abdelilah-Seyfried1,2,3 1Max Delbrück Center for Molecular Medicine, 2Institute of Biochemistry and Biology, University of Potsdam, 3Institute of Molecular Biology, Medizinische Hochschule Hannover To analyse cardiac gene expression profiles during zebrafish heart development, total RNA has to be extracted from isolated hearts. Here, we present a protocol for collecting functional/beating hearts by rapid manual dissection from zebrafish embryos to obtain cardiac-specific mRNA.