Immunohistochemistry: Basics and Methods (Paperback, 2010 ed.)

Combiningtwodifferentscienti?cdisciplines-morphologyandimmunochemistry- immunohistochemistryhasdevelopedasanimportantinstrumentinresearchand clinicalpathology. A basicunderstandingofunderlying principlesandpotential problemsisunavoidableifyouwanttobesuccessfulinyouruseofimmunohis- chemistry,aswellasingettingyourpaperspublishedandyourresearchgrants funded. Whilemanyexcellenttextsandmonographsexistwhichcovervariousaspects ofimmunohistochemistry,thelackofaconcisecomprehensiveguidetousing thesemethodswasamajormotivationforwritingthisbook. Ourintentionwasto createaneasy-to-readandfocusedresourcebasedonstate-of-the-artinformation forabroadaudiencerangingfromstudentsandtechnicalassistantstoexperienced researchers. Thishandbookhasaconciseformat,withprotocolsandinstructionsfor methodsimmediatelyfollowingtheshortintroductorytheoreticalmaterialineach chapter. BeingconsciousofthegrowingroleofInternetasaninformationsource, wehavefounditreasonableinmanycasestosubstitutecitingbooksandjournal publications with corresponding Internet websites. Where possible, commercial sourcesofreagents,kits,andequipmentarelistedthroughoutthetextinsteadof inaseparateindex. Thougheachchapterissmallandintroductory,thishandbook itself is self-suf?cient and provides a comprehensive look at the principles of immunohistochemistry. For readers wanting further depth of knowledge, each chapterisbackedupbyashortlistofcarefullyselectedoriginalarticles. Duringthelastdecade,pioneeringeffortsofhistochemistshaveledtoan- menseimprovementinthereagentsandprotocols. Theresearcherisurgedalwaysto determinethereasonforeverymethodandstepbeforedoingit. Thishandbookis intendedtohelpreaderstoavoidtroublesinthechoiceofanadequatemethod,which happenswhenusingstandardtextbooks. Forthishandbook,wecarefullyselected 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 colleaguesfromtheMunsterUniversityClinicforsharingprobesandreagents. IgorB. BuchwalowandWernerBocker Munster 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 ...13 2. 4 FluorophoreLabelsforFluorescenceMicroscopy ...15 2. 5 ColloidalGoldLabelsforElectronMicroscopy ...16 References ...17 3 ProbesProcessinginImmunohistochemistry ...19 3. 1 FixationinImmunohistochemistry ...19 3. 1. 1 FixationinAlcoholsandAcetone ...20 3. 1. 2 FixationinFormaldehyde ...20 3. 1. 3 EffectofFormaldehydeFixationon Antigen-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 ...13 2. 4 FluorophoreLabelsforFluorescenceMicroscopy ...15 2. 5 ColloidalGoldLabelsforElectronMicroscopy ...16 References ...17 3 ProbesProcessinginImmunohistochemistry ...19 3. 1 FixationinImmunohistochemistry ...19 3. 1. 1 FixationinAlcoholsandAcetone ...20 3. 1. 2 FixationinFormaldehyde ...20 3. 1. 3 EffectofFormaldehydeFixationon Antigen- immunohistochemistryhasdevelopedasanimportantinstrumentinresearchand clinicalpathology. A basicunderstandingofunderlying principlesandpotential problemsisunavoidableifyouwanttobesuccessfulinyouruseofimmunohis- chemistry,aswellasingettingyourpaperspublishedandyourresearchgrants funded. Whilemanyexcellenttextsandmonographsexistwhichcovervariousaspects ofimmunohistochemistry,thelackofaconcisecomprehensiveguidetousing thesemethodswasamajormotivationforwritingthisbook. Ourintentionwasto createaneasy-to-readandfocusedresourcebasedonstate-of-the-artinformation forabroadaudiencerangingfromstudentsandtechnicalassistantstoexperienced researchers. Thishandbookhasaconciseformat,withprotocolsandinstructionsfor methodsimmediatelyfollowingtheshortintroductorytheoreticalmaterialineach chapter. BeingconsciousofthegrowingroleofInternetasaninformationsource, wehavefounditreasonableinmanycasestosubstitutecitingbooksandjournal publications with corresponding Internet websites. Where possible, commercial sourcesofreagents,kits,andequipmentarelistedthroughoutthetextinsteadof inaseparateindex. Thougheachchapterissmallandintroductory,thishandbook itself is self-suf?cient and provides a comprehensive look at the principles of immunohistochemistry. For readers wanting further depth of knowledge, each chapterisbackedupbyashortlistofcarefullyselectedoriginalarticles. Duringthelastdecade,pioneeringeffortsofhistochemistshaveledtoan- menseimprovementinthereagentsandprotocols. Theresearcherisurgedalwaysto determinethereasonforeverymethodandstepbeforedoingit. Thishandbookis intendedtohelpreaderstoavoidtroublesinthechoiceofanadequatemethod,which happenswhenusingstandardtextbooks. Forthishandbook,wecarefullyselected 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 colleaguesfromtheMunsterUniversityClinicforsharingprobesandreagents. IgorB. BuchwalowandWernerBocker Munster 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 ...13 2. 4 FluorophoreLabelsforFluorescenceMicroscopy ...15 2. 5 ColloidalGoldLabelsforElectronMicroscopy ...16 References ...17 3 ProbesProcessinginImmunohistochemistry ...19 3. 1 FixationinImmunohistochemistry ...19 3. 1. 1 FixationinAlcoholsandAcetone ...20 3. 1. 2 FixationinFormaldehyde ...20 3. 1. 3 EffectofFormaldehydeFixationon Antigen-AntibodyBinding ...21 3. 2 Paraf?nSectionsforImmunohistochemicalAnalysis ...22 3. 2. 1 EmbeddingandCutting ...22 3. 2. 2 MountingParaf?nSectionsontoSlides ...23 3. 3 CryosectionsforImmunohistochemicalAnalysis ...24 3. 4 BuffersforWashingandAntibodyDilution ...25 3. 5 MountingFollowingImmunohistochemicalStaining ...27 3. 6 StorageFollowingImmunohistochemicalStaining ...28 References ...28 vii viii Contents 4 WorkingwithAntibodies ...31 4. 1 DirectImmunostainingMethod ...31 4. 2 IndirectImmunostainingMethod ...33 4. 3 TheChoiceofAntibodies ...35 4. 3. 1 TheChoiceofPrimaryAntibodies ...35 4. 3. 2 TheChoiceofSecondaryAntibodies ...

General

Imprint:	Springer-Verlag
Country of origin:	Germany
Release date:	October 2014
First published:	2010
Authors:	Igor B. Buchwalow • Werner Boecker
Dimensions:	235 x 155 x 9mm (L x W x T)
Format:	Paperback
Pages:	153
Edition:	2010 ed.
ISBN-13:	978-3-642-42502-8
Categories:	Books > Medicine > Pre-clinical medicine: basic sciences > Anatomy > Histology Books > Medicine > Clinical & internal medicine > Diseases & disorders > Immunology > General
LSN:	3-642-42502-X
Barcode:	9783642425028

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