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Animal Cell Electroporation and Electrofusion Protocols

Handbook / Manual

Series: Methods in Molecular Biology Volume: 48

Edited By: JA Nickloff

392 pages, 11 b/w photos, 46 line illus

Humana Press

Spiralbound | Dec 1995 | #43267 | ISBN: 089603304X
Availability: Usually dispatched within 1-2 weeks Details
NHBS Price: £108.00 $138/€129 approx

About this book

This superb collection of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells also includes proven animal cell electrofusion techniques for studies of somatic cell genetics and development, and for generating monoclonal antibodies. It is distinguished by its coverage of important model cell types from many organisms and tissue types and its detailed instructions for the growth and preparation of specific cells to achieve optimum transfection. Its extensive reference lists, citations of alternative transfer methods, advice on pitfalls to avoid, and descriptions of expected results ensure readily reproducible results even for beginners.

This volume should fulfill its aim, to take the guesswork out of experimental trials. - Society for General Microbiology Quarterly The book is very up-to-date, for instance it cover the most important recent developments in experiments with isolation of tumor suppressor genes in melanoma by cDNA microarray. The book represents an outstanding work, which should not be missed in all laboratories dealing with melanoma as well as in all departments where melanoma patients are diagnosed and treated. Those people who are interested to learn how the completion of the Human Genome Project significantly has been contributing to new progress in cancer management, exampled by human malignant melanoma, will certainly enjoy reading this publication as well. Neoplasma


Contents

Part I. Theory and Instrumentation. Electroporation Theory: Concepts and Mechanisms. Effects of Pulse Length and Strength on Electroporation Efficiency. Instrumentation. Part II. Electroporation Protocols. The Introduction of Proteins into Mammalian Cells by Electroporation. Electroporation of Antigen-Presenting Cells for T-Cell Recognition and Cytotoxic T-Lymphocyte Priming. Electroporation of Antibodies into Mammalian Cells. Electroporation of Adherent Cells In Situ for the Introduction of Nonpermeant Molecules. Electrotransformation of Chinese Hamster Ovary Cells. Electroporation of Rat Pituitary Cells. Electroporation of Plasmid DNA into Normal Human Fibroblasts. Electroporation-Mediated Gene Transfer into Hepatocytes. Electroporation of Human Lymphoblastoid Cells. The Use of Electroporated Bovine Spermatozoa to Transfer Foreign DNA into Oocytes. Electroporation of Embryonic Stem Cells for Generating Transgenic Mice and Studying In Vitro Differentiation. Electrotransfection with Intracellular Buffer. Effect of Cis-Located Human Satellite DNA on Electroporation Efficiency. Quantitation of Transient Gene Expression. Stable Integration of Vectors at High Copy Number for High-Level Expression in Animal Cells. Electroporation of Drosophila Embryos. Transformation of Fish Cells and Embryos. Electroporation of Cardiac Cells. Electroporation for Gene Therapy. Part III. Electrofusion Protocols. Electrofusion of Mammalian Cells. Stabilizing Antibody Secretion of Human Epstein Barr Virus-Activated B-Lymphocytes with Hybridoma Formation by Electrofusion. Electrofusion of Mammalian Oocytes and Embryonic Cells. Nuclear Transfer in Bovine Embryos. Electrofusion of Mouse Embryos to Produce Tetraploids. Spectrofluorometric Assay for Cell-Tissue Electrofusion. Cytometric Detection and Quantitation of Cell Cell Electrofusion Products. Index.

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