Immunohistochemistry and Immunocytochemistry

Essential Methods

Häftad, Engelska, 2017

Av Simon Renshaw, Simon Renshaw, UK) Renshaw, Simon (Abcam plc, Cambridge

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Immunohistochemistry and immunocytochemistry are invaluable tools for the visualization of tissue and cellular antigens in diagnostic and biological research environments. The need to obtain accurate, reliable and reproducible results is paramount.It is with this fundamental aim in mind that we have compiled Immunohistochemistry: Essential Methods. We have achieved this by examining each aspect of immunochemistry in turn, with each chapter including detailed information regarding the subject matter in question. Each chapter is written by an expert in their field and includes protocols that are typically used in their own research. Subjects covered are, amongst others, antibodies and their production; selection of reporter labels; immunochemical staining methods and experimental design (both using single and multiple reporter labels); quality assurance; automated immunochemistry; confocal microscopy and electron microscopy. In addition, benefits and limitations of each approach are discussed within the chapters.

Produktinformation

Utgivningsdatum2017-01-13
Mått170 x 244 x 13 mm
Vikt476 g
FormatHäftad
SpråkEngelska
SerieEssential Methods
Antal sidor256
Upplaga2
FörlagJohn Wiley and Sons Ltd
ISBN9781118717776

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Diagnostiska metoder inom Medicin

List of Contributors vii Preface ixAcknowledgements xiChapter 1: Antibodies for Immunochemistry 1Mark Cooper and Sheriden LummasIntroduction 1Immunogens for Antibody Production 5Antibody Production 12Antibody Purification 16Fragment Antibody Preparations 20Antibody Labelling 21Antibody Stability and Storage 23References 24Chapter 2: The Selection of Reporter Labels 25Judith LangenickIntroduction 25Enzymatic Labels 26Fluorescence Detection 29References 32Chapter 3: Immunohistochemistry and Immunocytochemistry 35Simon RenshawSpecimen Formats for Immunochemistry 36Fixation 37Processing Tissue Blocks to Paraffin Wax 46Microtomy 47Tissue Microarrays 47Specimen Storage 48Decalcification 49Antigen Retrieval 50Controls 56Immunochemical Staining Techniques (Optimizing a New Antibody) 57Counterstains 71Mounting 74Troubleshooting 76Examples of Immunostaining Photomicrographs 76Acknowledgements 101References 101Chapter 4: Multiple Immunochemical Staining Techniques 103Sofia KochIntroduction 103Methods and Approaches 115References 122Chapter 5: Quality Assurance in Immunochemistry 123Peter Jackson and Michael GandyIntroduction 123Methods and Approaches 125Automated Immunochemical Staining 147Troubleshooting 149References 154Chapter 6: Automated Immunochemistry 157Emanuel Schenck and Simon RenshawIntroduction 157Methods and Approaches 160Other Forms of Automation 164References 168Chapter 7: Confocal Microscopy 169Ann WheelerIntroduction 169When Should Confocal be Used? 173Applications: For Example Co-localization, Quantification, 3D Visualization and Kinetics 173How To Set Up a Confocal Experiment? 174References 198Further Readings 198Chapter 8: Ultrastructural Immunochemistry 199Jeremy Skepper and Janet PowellIntroduction 199Methods and Approaches 207References 222Index 227