
TheFluoroSpotassayisaninventivedevelopmentoftheELISpotassayandusesfluorescentdetectioninsteadofcolorimetric.Thisallowsforsimultaneousdetectionofseveralanalytes.ThehumanIgG/IgA/IgMB-cellFluoroSpotkitenablesdetectionofIgG-,IgA-andIgM-secretingcellsinthesamewell.
Thekitcontainsmatchedquantitiesofcaptureantibodiesandfluorophore-conjugateddetectionantibodies,lowfluorescentPVDFplatesandafluorescenceenhancer.ThepolyclonalactivatorsR848andrecombinanthumanIL-2areincludedforpre-stimulationofmemoryBcells.
Foranalysis,werecommendtheuseofanautomatedFluoroSpotreaderequippedwithfiltersforFITC(excitation490nm/emission510nm),Cy3(excitation550nm/emission570nm)andCy5(excitation640nm/emission660nm).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.
IgM
MonomericIgM(ImmunoglobulinM)isfoundonthesurfaceofBcellswhereastheIgMmoleculessecretedfromplasmaBcellsarepentamers,i.e.fiveIgMmoleculesjoinedtogetherbytheJchain.IgMconstitutesapproximately10%ofthetotalIgconcentrationinserum.IgMisthefirstantibodyisotypeinducedinatypicalimmuneresponseandhenceistheisotypewiththelowestaffinityforantigens.Duetoitsstructure,with10antigen-bindingsitesperIgMpentamer,itcanneverthelessbinde.g.surface-boundantigenswithaveryhighAvidity.
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Size: | 10plates |
Otherformats: | ClicktoviewallHumanIgA,IgG,IgMproducts |
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尤其是获得SCI文献引用的产品,
绝对ok的
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。

