TheFluoroSpotassayisaninventivedevelopmentoftheELISpotassayandusesfluorescentdetectioninsteadofcolorimetric.Thisallowsforsimultaneousdetectionofseveralanalytes.ThemonkeyIgG/IgAB-cellFluoroSpotkitenablesdetectionofIgG-andIgA-secretingcellsinthesamewell.

Thekitcontainsmatchedquantitiesofcaptureantibodiesandfluorophore-conjugateddetectionantibodies,lowfluorescentPVDFplatesandafluorescenceenhancer.ThepolyclonalactivatorsR848andrecombinanthumanIL-2areincludedforpre-stimulationofmemoryBcells.

TheantibodiesinthiskitreactwithhumanIgGandIgAandcross-reactwithIgGandIgAfromnon-humanprimates.

Foranalysis,werecommendtheuseofanautomatedFluoroSpotreaderequippedwithfiltersforFITC(excitation490nm/emission510nm)andCy3(excitation550nm/emission570nm).Filtersshouldhavehighspecificitytoavoidbleed-throughartifacts.

AnalyteInformation

IgA
IgA(ImmunoglobulinA)representsonly10-15%ofthetotalIgmoleculesinserumbutisthepredominatingIgisotypeinsaliva,tears,breastmilkandinthegastrointestinalandurogenitaltract.Inserum,theIgAmoleculeiscommonlyfoundasamonomerofeitherIgA1orIgA2subclassbutcanalsoexistasdimersorlargerpolymericforms.TheIgAfoundinsecretions(secretoryIgAorsIgA)iscommonlyadimerwithtwoIgAmoleculesjoinedbytheJchain.SecretoryIgAisthemostimportantIgisotypeinmucoussurfaces,theareaofentryformostpathogens.

IgG
IgG(ImmunoglobulinG)isthemostabundantIgisotypeinserumandmakesupapproximately80%ofallserumImmunoglobulins.Inhumans,therearefoursubclassesofIgGwithhighestserumconcentrationsofIgG1followedbyIgG2,IgG3andIgG4.InmicetheIgGsubclassesaredefinedIgG1,IgG2a/c,IgG2bandIgG3.TheIgGmoleculeconsistsoftwoheavyandtwolightschains(κorλ)resultinginamoleculewithtwoarmsforantigenbinding.HighlevelsofIgGantibodiesareinducedfollowingtheinitialIgMresponseinatypicalimmuneresponsetoantigens.

   Mabtech 公司于1986年创办于瑞典斯德哥尔摩大学免疫学系，是一家欧洲的集研究，开发，销售，技术支持为一体的高科技免疫生物技术公司。Mabtech公司始终专业致力于研发新的单克隆抗体，生产创新的免疫试剂和销售高品质的全配套的免疫试剂盒。公司核心业务是致力于优化发展酶联免疫斑点法技术 (ELISpot)、荧光免疫斑点法技术 (FluoroSpot) 和酶联免疫吸附试验技术 (ELISA) 等免疫学方法。Mabtech公司的产品质量体系符合ISO 9001和ISO 13485国际标准。