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008			100301s2010 gw | s |||| 0|eng d
020			_a9783642046094 _9978-3-642-04609-4
040			_cMX-MeUAM
050		4	_aQR180-189.5
082	0	4	_a616.079 _223
100	1		_aBuchwalow, Igor B. _eauthor.
245	1	0	_aImmunohistochemistry: Basics and Methods _h[recurso electrónico] / _cby Igor B. Buchwalow, Werner Böcker.
264		1	_aBerlin, Heidelberg : _bSpringer Berlin Heidelberg, _c2010.
300			_aX, 153 p. _bonline resource.
336			_atext _btxt _2rdacontent
337			_acomputer _bc _2rdamedia
338			_aonline resource _bcr _2rdacarrier
347			_atext file _bPDF _2rda
505	0		_aAntibodies for Immunohistochemistry -- Antibody Labeling and the Choice of the Label -- Probes Processing in Immunohistochemistry -- Working with Antibodies -- Background Staining, Autofluorescence and Blocking Steps -- Immunostaining Enhancement -- Multiple Multicolor Immunoenzyme Staining -- Multiple Immunofluorescence Staining -- Antigen Detection on Tissues Using Primary Antibody Raised in the Same Species -- Probes for Staining Specific Cellular Organelles -- The Use of Epitope Tags in Histochemistry -- Immunohistochemistry at the Ultrastructural Level -- Diagnostic Immunohistochemistry -- A Picture Is Worth a Thousand Words.
520			_aCombiningtwodifferentscienti?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 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 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 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 . . . . . . . . . . . . . . . . . . . . . . . . .
650		0	_aMedicine.
650		0	_aImmunology.
650		0	_aMonoclonal antibodies.
650		0	_aMedical laboratories.
650		0	_aCytology.
650		0	_aCytology _xResearch_xMethodology.
650	1	4	_aBiomedicine.
650	2	4	_aImmunology.
650	2	4	_aBiological Techniques.
650	2	4	_aCell Biology.
650	2	4	_aLaboratory Medicine.
650	2	4	_aAntibodies.
700	1		_aBöcker, Werner. _eauthor.
710	2		_aSpringerLink (Online service)
773	0		_tSpringer eBooks
776	0	8	_iPrinted edition: _z9783642046087
856	4	0	_zLibro electrónico _uhttp://148.231.10.114:2048/login?url=http://link.springer.com/book/10.1007/978-3-642-04609-4
596			_a19
942			_cLIBRO_ELEC
999			_c201543 _d201543