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Methods in Cell Biology PDF[1]

Methods in Cell Biology PDF[1]

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Published by azabuzi
this will help who are interested in experimental cell and molecular biology. all the contents are laboratory oriented based on basic experimental procedure.
this will help who are interested in experimental cell and molecular biology. all the contents are laboratory oriented based on basic experimental procedure.

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Published by: azabuzi on Feb 13, 2010
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12/12/2012

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1
Lecture Notes forMethods in Cell Biology
(TRMD 623)Instructor:Mark F. Wiser
 
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TABLE OF CONTENTS
 PREFACE—COURSE DESCRIPTION................................................................................7CHAPTER 1--MICROSCOPY..............................................................................................9LIGHT MICROSCOPY.............................................................................................9ELECTRON MICROSCOPY...................................................................................12MICROSCOPY APPENDIX. ALDEHYDE FIXATIVES......................................16CHAPTER 2--SPECTROPHOTOMETRY..........................................................................19SPECTROPHOTOMETRY THEORY....................................................................19INSTRUMENTATION............................................................................................21APPLICATIONS OF SPECTROPHOTOMETRY..................................................21FACTORS AFFECTING ABSORPTION...............................................................22VARIATIONS IN SPECTROPHOTOMETRY.......................................................22APPENDIX I. PHOTOMULTIPLIER TUBE..........................................................24APPENDIX II. CALCULATION OF ENZYME ACTIVITY.................................25CHAPTER 3--FLUORESCENCE........................................................................................29ENZYME ASSAYS AND FLUOROMETERS.......................................................29FLUORESCENCE MICROSCOPY.........................................................................30FLOW CYTOMETRY.............................................................................................33CHAPTER 4--RADIOCHEMISTRY...................................................................................35ATOMIC STRUCTURE AND RADIOACTIVE DECAY......................................35PROPERTIES OF RADIOISOTOPES....................................................................36MEASUREMENT OF ß-RADIATION...................................................................37GAMMA-RAY DETECTION.................................................................................42SAFETY...................................................................................................................42APPENDIX 1. INTERNAL STANDARDS.............................................................43APPENDIX 2. CHANNELS RATIO.......................................................................44APPENDIX 3. MULTIPLE ISOTOPES..................................................................45CHAPTER 5--pH AND BUFFERS......................................................................................47DEFINITIONS.........................................................................................................47HENDERSON-HASSELBACH...............................................................................47INSTRUMENTATION............................................................................................48COMPLICATIONS..................................................................................................48BUFFERING CAPACITY.......................................................................................49BUFFER SELECTION............................................................................................49BUFFER APPENDIX..............................................................................................50
 
3CHAPTER 6--CENTRIFUGATION....................................................................................53SEDIMENTATION THEORY.................................................................................53PREPARATIVE CENTRIFUGATION...................................................................55CENTRIFUGATION APPENDIX 1. RCF CALCULATION.................................58CHAPTER 7--INTRODUCTION TO PROTEINS..............................................................61PROTEIN STRUCTURE.........................................................................................61PROTEIN STABILITY............................................................................................63PROTEIN ASSAYS.................................................................................................64APPENDIX. AMINO ACIDS: PROPERTIES AND STRUCTURE......................66CHAPTER 8--DIFFERENTIAL PRECIPITATION OF PROTEINS..................................69PROCEDURE FOR (NH
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PRECIPITATION.................................................69OTHER PROTEIN PRECIPITATION METHODS................................................70APPENDIX. AMMONIUM SULFATE NOMOGRAM.........................................71CHAPTER 9--CHROMATOGRAPHY...............................................................................73BASIC PRINCIPALS...............................................................................................73EQUIPMENT...........................................................................................................74ADSORPTION CHROMATOGRAPHY.................................................................75ION EXCHANGE CHROMATOGRAPHY............................................................75HYDROPHOBIC CHROMATOGRAPHY.............................................................79Polar Solvent.........................................................................................................................80GEL FILTRATION CHROMATOGRAPHY..........................................................80AFFINITY CHROMATOGRAPHY........................................................................81CHAPTER 10--MEMBRANES AND DETERGENTS.......................................................83TYPES OF MEMBRANE PROTEINS....................................................................84DETERGENTS.........................................................................................................85DETERGENT REMOVAL......................................................................................87APPENDIX 1. CELL DISRUPTION TECHNIQUES.............................................88CHAPTER 11--ELECTROPHORESIS................................................................................91GEL ELECTROPHORESIS.....................................................................................91EQUIPMENT...........................................................................................................93SDS-PAGE...............................................................................................................94ISOELECTRIC FOCUSING....................................................................................96TWO-DIMENSIONAL GEL ELECTROPHORESIS..............................................97PROTEIN DETECTION FOLLOWING ELECTROPHORESIS............................98PREPARATIVE ELECTROPHORESIS...............................................................101ELECTROPHORESIS APPENDIX 1. SIZE CALCULATION............................104

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