Immunohistochemistry: Basics and Methods

establishedmethodsandeasy-to-adoptprotocols,payingattentiontomoderndev- opmentsinimmunohistochemistry,suchasantigenretrieval,signalampli?cation, the use of epitope tags in immunohistochemistry, multiple immunolabeling or diagnosticimmunohistochemistry. Eachofthemethodsdescribedinthishandbook v vi Preface was provedby the authors; many of these methods are routinely used in daily practiceintheirinstitute. Allthepracticalmethodsadvocatedareclearlydescribed, withaccompanyingtables,andtheresultsobtainableareillustratedwithcolour micrographs. Acknowledgements We thank Vera Samoilova forthe perfect technical assistance and other colleaguesfromtheMu"nsterUniversityClinicforsharingprobesandreagents. IgorB. BuchwalowandWernerBo"cker Mu"nster Contents 1 AntibodiesforImmunohistochemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1. 1 StructureofAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1. 2 PolyclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1. 3 MouseMonoclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1. 4 RabbitMonoclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1. 5 ProteinAandProteinGinImmunohistochemistry . . . . . . . . . . . . . . . . . . . 7 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 2 AntibodyLabelingandtheChoiceoftheLabel . . . . . . . . . . . . . . . . . . . . . . . . 9 2. 1 CovalentLabelingofAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 2. 2 Non-CovalentLabelingofPrimaryAntibodieswith LabeledFabFragments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 2. 3 EnzymeLabelsforLightMicroscopy . . . . . . . . . .