TheFluoroSpotassayisaninventivedevelopmentoftheELISpotassayandusesfluorescentdetectioninsteadofcolorimetric.Thisallowsforsimultaneousdetectionofseveralanalytes,whichisamajoradvantage.ThehumanIL-1β/GM-CSFFluoroSpotkitenablesconvenientenumerationofcellssecretingeitheroneorbothanalytes.Thekitcontainslowfluorescentplates,matchedquantitiesofmonoclonalcaptureanddetectionantibodies,secondaryreagentsconjugatedtofluorophoresandafluorescenceenhancer.LPSisincluded.
Theassayisrecommendedfortheanalysisofmacrophages,monocytesanddendriticcells.ThesekitshavenotbeenevaluatedforT-cells.
Foranalysis,werecommendtheuseofanautomatedFluoroSpotreaderequippedwithfiltersforFITC(excitation490nm/emission510nm)andCy3(excitation550nm/emission570nm).Filtersshouldhavehighspecificitytoavoidbleed-throughartifacts.Doublesecretingcellsareidentifiedbyacomputerizedoverlay.
AnalyteInformation
GM-CSF
Granulocytemacrophage-colonystimulatingfactor(GM-CSF)canbeproducedbyseveralcelltypesamongothersactivatedTcells,macrophages,endothelialcellsandstromalcells.GM-CSFstimulatesthesurvivalandfunctionalactivitiesofneutrophils,eosinophils,andmonocyte-macrophages.ItalsostimulatesthedifferentiationandproliferationofhematologicalProgenitors.
IL-1β
Interleukin-1ß(IL-1ßorIL-1F2,alsoknownasInterleukin-1beta,IL-1beta,IL1bandInterleukin-1beta)isaproinflammatorycytokineandinducerofacutephaseresponses.IL-1ßisproducedprimarilybymonocytes,macrophages,anddendriticcellsafterinductionbymicrobes.
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| Size: | 10plates |
| Otherformats: | ClicktoviewallHumanGM-CSF,IL-1βproducts |
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用Mabteck公司的ALP标记抗体吧，配合Millipore公司的Multiscreen-IP96孔板，所有的试剂都可以减半，照样可以出非常好的结果，4*96孔的试剂盒我一直做8*96孔，但BCIP/NBT显色液没试过减半，因为它很便宜。记得当年第一次做实验就出了很好的结果，经过不断摸索，做出的蓝紫色斑点确实非常漂亮，几乎没有非特异斑点，嗯，ELISPOT是我最拿手的实验之一。
关于PBMC:
拿到血液标本后，首先我要做的是尽快分离PBMC（一般2h内），国产的分离液里天津灏洋的比较好，然后用冻存液冻存PBMC。PBMC的冻存过程比一般的细胞株要更为严格，最好用冻存器。ELISPOT试验的第一天，在包被抗体的同时，将细胞复苏，完全培养液培养过夜，第二天收集细胞并计数即可。
尤其是获得SCI文献引用的产品，
绝对ok的
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！！！
