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Laboratory Exercises and Techniques in Cellular Biology
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Introduction vii Acknowledgment ix 1 Introduction and Microscopy 1 (Karp chapter 1 & 18) 2 Enzyme Kinetics 11 (Karp chapter 3 & 11) 3 Membrane Permeability 19 (Karp chapter 4) 4 Protein Transport to the Plasma Membrane 27 (Karp chapter 4 & 8) 5 Cell Fractionation 33 (Karp chapter 5 & 12) 6 Isolation and Activity of Mitochondria 41 (Karp chapter 3 & 5) 7 Staining of the Extracellular Space 49 (Karp chapter 7) 8 Protoplast Isolation and Vacuole Staining 57 (Karp chapter 8) 9 Isolation of Vacuoles 65 (Karp chapter 8) 10 Staining of Autophagosomes and Lysosomes 71 (Karp chapter 8) 11 Staining of the Cytoskeleton 77 (Karp chapter 9) 12 Cell Motility 85 (Karp chapter 9) 13 Staining of the Nucleus and Nucleic Acids 95 (Karp chapter 12) 14 Staining of Chromosome and Cell Cycle Analysis 105 (Karp chapter 14) 15 Staining for Cell Viability and Apoptosis 113 (Karp chapter 15 & 16) 16 Staining of Immune Cells 121 (Karp chapter 17) 17 Protein Isolation, Fractionation, and SDS-PAGE 131 (Karp chapter 18) 18 Genomic DNA Isolation, Purification, and the Polymerase Chain Reaction 139 (Karp chapter 18) 19 Ligation and Cloning 145 (Karp chapter 18) 20 Restriction Fragment Length Polymorphism Analysis and Agarose Gel Electrophoresis 151 (Karp chapter 10 & 18) 21 Staining of Peroxisomes* 157 (Karp chapter 5) 22 Short-Term Aggregation Culture* 163 (Karp chapter 7) 23 Staining of the Endoplasmic Reticulum and the Golgi Apparatus* 171 (Karp chapter 8) 24 The Endocytic Pathway (FM4-64 Staining)* 179 (Karp chapter 4 & 8) 25 Detection of MAP Kinase Signaling* 187 (Karp chapter 15) 26 Cancer* 193 (Karp chapter 16) 27 Western Blot* 197 (Karp chapter 18) 28 Enzyme-Linked Immunosorbent Assay* 203 (Karp chapter 18) 29 Southern and Northern Blotting* 211 (Karp chapter 18) 30 Using Online Research Tools* 217 (Karp chapter 10 & 11) Appendix A: A Guide to Staining Techniques & Photomicroscopy (Available online at www.wiley.com/college/contento) Appendix B: Careers in Cell Biology (Available online at www.wiley.com/college/contento) Instructor?s Notes (Available online at www.wiley.com/college/contento) Index 223

About the Author

Anthony Contento is a plant scientist and lecturer at Iowa State University. His research interests include plant responses to abiotic stresses, macroautophagy, prion biochemistry and translational medicine. He earned his BS at SUNY Fredonia in Recombinant Gene Technology in 1995, with minors in Chemistry, English, and History. He earned his Ph.D in Molecular Biology at the University of Wyoming in 2001.

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