Red blood cell-mediated microinjection: methodological considerations |
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| Authors: | P A Netland J F Dice |
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| Affiliation: | 1. Department of Anesthesiology, University of Rochester Medical Center, Rochester, NY, USA;2. Department of Chemistry, University of Rochester, Rochester, NY, USA;1. The State Key Laboratory of Medical Molecular Biology, Neuroscience Center, Medical Primates Research Center and Department of Molecular Biology and Biochemistry, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China;2. Department of Anatomy and Histology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China;3. Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118, China;4. Department of Molecular, Cellular, and Developmental Biology, Yale University, P.O. Box 208103, New Haven, CT 06520, USA;1. Department of Materials Engineering, Indian Institute of Science, Bangalore 560012, India;2. Department of Medical Devices, National Institute of Pharmaceutical Education and Research, Ahmedabad, Gandhinagar, 382355, Gujarat, India;3. Smart Materials and Surfaces Laboratory, Faculty of Engineering and Environment, Northumbria University, Newcastle upon Tyne, NE1 8ST, United Kingdom;1. Marseille Cancer Research Center (CRCM), U1068 INSERM, UMR7258 CNRS, Aix Marseille University, Institut Paoli-Calmettes (Equipe labellisée Ligue), Marseille 13009, France;2. Department of Biology, University of Copenhagen, 2200 Copenhagen N, Denmark;1. National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;2. State Key Laboratory of Virology and Center for Analytical Microbiology, Wuhan Institute of Virology, Chinese Academy of Sciences, China;1. Laboratory of Advanced Polymers, Institute of Chemistry, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves, 9500, Lab K-208 A, Porto Alegre, RS CEP 91501-970, Brazil;2. Laboratory of Fungi Importance Medicine and Biotechnology, Institute of Biotechnology, Universidade Federal do Rio Grande do Sul, Lab 220, Av. Bento Gonçalves, 9500, Porto Alegre, RS CEP 91501-970, Brazil;3. Post-Graduate Program of Materials Science, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves, 9500, Sala A216, Porto Alegre, RS CEP 91501-970, Brazil |
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| Abstract: | ![]()
Red blood cell-mediated microinjection is a powerful approach to introducing proteins into the cytoplasm of cultured cells. In the course of our microinjection studies of intracellular protein degradation, we have encountered several potential problems with certain proteins. The microinjection procedure may be accompanied by denaturation of protein by radiolabeling procedures, binding of protein to red cell ghosts during loading, degradation of protein by the red cell ghost prior to microinjection, and adsorption of protein that leaks from red cell ghosts in the presence of fusogen to the fibroblast monolayer. We conclude with a list of points that must be considered prior to use of red cell-mediated microinjection to study a particular protein. |
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