资源描述：
《elispot技术原理及实验方法介绍》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、ELISPOT技术原理及实验方法介绍IntroductionofELISPOTtechnologyprincipleandexperimentmethodClicknumber:590releasedate:2009-12-30source:thissiteisforreferenceonly,declinedtoreprint,otherwiseresponsibleELISPOTtechnologyprincipleWiththewidespreadapplicationofenzyme-linkedimmunosorbentassayinmedicalandbiologicalfie
2、lds,newbreakthroughshavebeenmadeinthedetectionofvariouscytokinesandantibodiesinvitro.Intheresearchonthemechanismofimmuneresponsewhenusedenzyme-linkedimmunosorbentassay(ELISA)freecytokinedetectioninbodyfluids(CK)orantibody,butbecauseofthedifferenceoffreecirculatingantibodyorCKhalfmourningperiod,whi
3、chiscontinuouslymetabolisminbodyfluidsorincombinationwithtargetorgans,butnotexactlyreflecttheinvivothelevelofantibodyandCK.In80s,foreignresearchersestablishedsolidphaseenzyme-linkedimmunosorbentassay(ELISPOT)todetectspecificantibodysecretingcellsandCKsecretingcellsinvitroaccordingtothebasicprincip
4、leofELISAtechnology.Becauseofitshighspecificityandsensitivity,itiswidelyusedathomeandabroad・Itisofgreatsignificancetoexplorethepathogenesisofautoimmunediseases・ELISPOTisfromELISA,andabreakthroughofthetraditionalELISAmethodisanextensionofquantitativeELISAtechnologyandnewdevelopment・Botharetestsforc
5、ytokinesorothersolubleproteinsproducedbycells,thebiggestdifferencebetweenthemisthat:1,ELISA,theabsorbancewasmeasuredontheenzymescalebycolorreaction,andthetotalamountofsolubleproteinwasobtainedbycomparingwiththestandardcurve・2,ELISPOTbycolorreaction,showingclearspotsinthecorrespondingpositionofthes
6、ecretionofsolubleprotein,canbedirectlyunderthemicroscopecountingspotsorthroughtheELISPOTanalysissystemforcountingspots,1spotsrepresenting1cells,tocalculatethefrequencyofcellssecretetheprotein・(somestudiesnotonlymeasurecytokineproduction,butalsodetectcellfrequenciesthatsecretethiscytokine)3,because
7、ofthedetectionofsinglecelllevels,ELISPOTismoresensitivethanELISAandlimiteddilution,andcandetect1secretedproteinsfrom200thousand-30millioncells・4,captureantibodiesforBD,R&D,Mabtachproductionofhighaffinity,highspec
