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.
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Otherformats: | ClicktoviewallNHPIgA,IgGproducts |
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请问一般的实验室可以进行ELIspot吗?(我可以买到现成的试剂盒,资金这个应该没有问题。)ELIspot的难点在哪里?多谢大虾们的指点!这个对我很重要,多谢!!!
我想用酶联免疫斑点(ELISPOT)技术测定NK细胞活性,但不知道哪里能买到试剂盒,特向大家求助,谢谢。