Deprecated: Creation of dynamic property cls_session::$session_data_table is deprecated in /www/sites/www.188bio.com/index/systems/cls_session.php on line 49
Antibodies-Online/Rat Cytokine Array Q1/ABIN625800/8 samples188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
您好,欢迎您进入188进口试剂采购网网站! 服务热线:4000-520-616
蚂蚁淘商城 | 现货促销 | 科研狗 | 生物在线
产品资料

Antibodies-Online/Rat Cytokine Array Q1/ABIN625800/8 samples

Reactivity
Rat(Rattus)
MethodType
SandwichELISA
Application
AntibodyArray(AA),MultiplexELISA(mpELISA)
Options
Bulkdiscount
Supplier
SupplierProductNo.
PurposeQuantibody®RatCytokineArray1Kit.Detects10RatCytokines.Suitableforallliquidsampletypes.
BrandQuantibody®
SampleTypeBodyFluids,CellCultureSupernatant,Plasma,Serum,CellLysate,TissueLysate
AnalyticalMethodQuantitative
DetectionMethodFluorometric
SpecificityCINC-2,CINC-3,CNTF,Fractalkine(CX3CL1),GM-CSF,IL-1alpha(IL-1F1),IL-4,LIX,beta-NGF,VEGF-A
Characteristics
  • RunninganarrayislikerunningdozensofELISAssimultaneously.
  • Quantibodyarraysarestunninglysimpletouse,read,andanalyze.
  • Eachpanelcanquantifyupto40differentbioMarkerssimultaneously,andindividualpanelscanbemultiplexedtoquantifyasmanyas660differentbiomarkersatonetime.
  • Theentireprocesscanbecompletedinjust4-6hours.
  • Morecost-effectivethantrADItionalELISA
  • Highspecificityandsystemreproducibility
  • Suitablefordiversesampletypes
  • Lowsamplevolumerequirement:50μLorless
  • Well-suitedforhighthroughputassays

    • Morecost-effectivethantraditionalELISA
    • Highspecificityandsystemreproducibility
    • Suitablefordiversesampletypes
    • Lowsamplevolumerequirement:50μLorless
    • Getresultssameday(6-hourprocessingtime)
    • Well-suitedforhighthroughputassays
    • QAnalyzersoftwareprovidesone-stepcomputation
ComponentsGlassChipwithantibodyarrays
SampleDiluent
Lyophilizedproteinstandardmix
Detectionantibodycocktail
Streptavidin-Fluorescentdye
Washbuffer
MaterialnotincludedDistilledordeionizedwater
Smallplasticboxesorcontainers
Pipettors,Pipettetipsandothercommonlabconsumables
Orbitalshakeroroscillatingrocker
Aluminumfoil
Genemicroarrayscannerorsimilarlaserfluorescencescanner
BackgroundCytokinesplayanimportantroleininnateimmunity,apoptosis,angiogenesis,cellgrowthanddifferentiation.Theyareinvolvedininteractionsbetweendifferentcelltypes,cellularresponsestoenvironmentalconditions,andmaintenanceofhomeostasis.Inaddition,cytokinesarealsoinvolvedinmostdiseaseprocesses,includingcancerandcardiacdiseases.
ApplicationNotesCompletelycoverarrayareawithsampleorbufferduringincubation.Avoidfoamingduringincubationsteps.Performallincubationandwashstepsundergentlerockingorrotation.Covertheincubationchamberwithadhesivefilmduringincubation,particularlywhenincubationismorethan2hoursor<70 μLofsampleorreagentisused.Severalincubationstepssuchasstep6(blocking),step7(sampleincubation),step10(detectionantibodyincubation),orstep13(Cy3equivalentdyestreptavidinincubation)maybedoneovernightat4 °C.Pleasemakesuretocovertheincubationchambertightlytopreventevaporation.
Comment

TheQuantibodyarraysarequantitativemultiplexELISAarraysfeaturingfluorescentdetection.Theantibodiesarespottedonglassslidesolidsupportsandrequirealaserscannerfordatacollection.Cytokinestandardsareprovidedwiththearrayforcalculationoftargetproteinconcentrations.
AllQuantibodyarraysfeaturethesandwichimmunoassayprinciple,utilizinganimmobilizedcaptureantibodyalongwithacorrespondingbiotinylateddetectionantibody.

SampleVolume100μL
AssayTime6h
PlateGlassSlide
Protocol
  1. EachQuantibodyarraystartswithasingleglassmicroscopeslide,whichactsasasupportforthearray.Slidesaresegmentedusingarubbergasket.Upto8samplesmayassayedusingasingleslide.
  2. Antibodiesagainstavarietyofdifferentantigens(upto40biomarkersperslide)areprintedontotheglassslide.Replicatesareincluded,savingyoubothtimeandprecioussamplevolume.
  3. Theend-useraddseitherknownconcentrationstandards(included)oraqueoussampletoeachwellontheslide.Antibodiesontheslidecaptureantigenofffromthesampleorstandard.
  4. Theend-useraddsadetectionmixcontainingpairedantibodies(compatIBLewiththeprimariespre-coatedontheslide)conjugatedtoafluorescentdyefordetection.
  5. Fluorescentsignalfromeachspotisreadusingalaserslidescanner.Theintensityfromeachspotiscomparedtothestandardcurve,andaquantitativeexpressionprofileforrelevantbiomarkersisestablished.
SamplePreparation

Useserum-freeconditionedmediaifpossible.Ifserum-containingconditionedmediaisrequired,itishighlyrecommendedthatcompletemediumbeusedasacontrolsincemanytypesofseracontainscytokines.Werecommendthefollowingparametersforyoursamples:50to100loforiginalordilutedserum,plasma,cellculturemedia,orotherbodyfluid,or50-500g/mlofproteinforcellandtissuelysates.Ifyouexperiencehighbackgroundorthereadingsexceedthedetectionrange,furtherdilutionofyoursampleisrecommended.

AssayProcedure
  1. Takeouttheglassslidefromthebox,andletitequilibratetoroomtemperatureinsidethesealedplasticbagfor20-30minutes.Removeslidefromtheplasticbag,peeloffthecoverfilm,andletitairdryforanother1-2hours.
    2.ReconstitutetheCytokineStandardMix(lyophilized)byadding500µlSampleDiluenttothetube.Forbestrecovery,alwaysquick-spinvialpriortoopening.Dissolvethepowderthoroughlybyagentlemix.LabeledthetubeasStd1.
    3.Label6cleanmicrocentrifugetubesasStd2toStd7.Add200µlSampleDiluenttoeachofthetubes.
    4.Pipette100µlStd1intotubeStd2andmixgently.Perform5moreserialdilutionsbyadding100µlStd2totubeStd3andsoon.
    5.Add100µlSampleDiluenttoanothertubelabeledasCNTRL.DonotaddstandardcytokinesorsamplestotheCNTRLtube,whichwillbeusedasnegativecontrol.Forbestresults,includeasetofstandardsineachslide.
    6.Add100µlSampleDiluentintoeachwellandincubateatroomtemperaturefor30minutestoblockslides.
    7.Decantbufferfromeachwell.Add100µlstandardcytokinesorsamplestoeachwell.Incubatearraysatroomtemperaturefor1-2hour.
    8.Wash:
    -Decantthesamplesfromeachwell,andwash5times(5mineach)with150µlof1XWashBufferIatroomtemperaturewithgentlerocking.Completelyremovewashbufferineachwashstep.Dilute20xWashBufferIwithH2O.
    -Decantthe1xWashBufferIfromeachwell,wash2times(5mineach)with150µlof1XWashBufferIIatroomtemperaturewithgentlerocking.Completelyremovewashbufferineachwashstep.Dilute20XWashBufferIIwithH2O.
    9.Reconstitutethedetectionantibodybyadding1.4mlofSampleDiluenttothetube.Spinbriefly.
    10.Add80µlofthedetectionantibodycocktailtoeachwell.Incubateatroomtemperaturefor1-2hour.
    11.Decantthesamplesfromeachwell,andwash5times(5minseach)with150µlof1XWashBufferIandthen2timeswith150µlof1xWashBufferIIatroomtemperaturewithgentlerocking.Completelyremovewashbufferineachwashstep.
    12.Afterbrieflyspinningdown,add1.4mlofSampleDiluenttoCy3equivalentdye-conjugatedstreptavidintube.Mixgently.
    13.Add80µlofCy3equivalentdye-conjugatedstreptavidintoeachwell.Coverthedevicewithaluminumfoiltoavoidexposuretolightorincubateindarkroom.Incubateatroomtemperaturefor1hour.
    14.Decantthesamplesfromeachwell,andwash5times(5minseach)with150µlof1XWashBufferIatroomtemperaturewithgentlerocking.Completelyremovewashbufferineachwashstep.
    15.Disassemblethedevicebypushingclipsoutwardfromtheslideside.Carefullyremovetheslidefromthegasket.
    16.PlacetheslideintheSlideWasher/Dryer(a4-slideholder/centrifugetube),addenough1xWashBufferI(about30ml)tocoverthewholeslide,andthengentlyshakeatroomtemperaturefor15minutes.DecantWashBufferI.Washwith1xWashBufferII(about30ml)andgentlyshakeatroomtemperaturefor5minutes.
    17.Removewaterdropletscompletelybygentlyapplyingsuctionwithapipettetoremovewaterdroplets.Donottouchthearray,onlythesides.
    18.Imaging:ThesignalscanbevisualizedthroughuseofalaserscannerequippedwithaCy3wavelength(greenchannel)suchasAxonGenePix.Makesurethatthesignalfromthewellcontainingthehigheststandardconcentration(Std1)receivesthehighestpossiblereading,yetremainsunsaturated.
CalculationofResults

DataextractioncanbedoneusingtheGALfilethatisspecificforthisarrayalongwiththemicroarrayanalysissoftware(GenePix,ScanArrayExpress,ArrayVision,MicroVigene,etc.).

AssayPrecisionReproducibility:CV< 20%
RestrictionsForResearchUseonly
HandlingAdviceDonottouchthesurfaceoftheslides,asthemicroarrayslidesareverysensitive.Holdtheslidesbytheedgesonly.Handleallbuffersandslideswithpowderfreegloves.Handleglassslide/sincleanenvironment.TheQuantibodyslidesdonothavebarcodes.Tohelpdistinguishoneslidefromanother,transcribetheslideserialnumberfromtheslidebagtothebackoftheslidewithanultra-finepointpermanentmarker.PleaseNote:Redpermanentmarkercansignificantlyinterferewithfluorescentsignaldetection.Werecommendmarkingyourslideswithagreen,blueorblackultra-finepointpermanentmarker.Pleasewritethenumberontheverybottomedgeoftheslide.Donotwriteonthearrayedwellareas.
Storage-20°C
StorageCommentForbestresults,storetheentirekitfrozenat-20°Cuponarrival.Storedfrozen,thekitwillbestableforatleast6monthswhichisthedurationoftheproductwarrantyperiod.Oncethawed,storearrayslide(s),standardmix,detectionantibodycocktail,andCy3-ConjugatedStreptavidinat-20°Candallotherreagentsundilutedat4°Cfornomorethan3months.
ExpiryDate6months
SupplierImages
 image for Rat Cytokine Array Q1 (ABIN625800)RatCytokineArrayQ1
Productcitedin:Meireles,Marques,Norberto,Fernandes,Mateus,Rendeiro,Spencer,Faria,Calhau:"Theimpactofchronicblackberryintakeontheneuroinflammatorystatusofratsfedastandardorhigh-fatdiet."in:TheJournalofnutritionalbiochemistry,Vol.26,Issue11,pp.1166-73,2015(PubMed).

Hanaoka,Nicolls,Fontenot,Kraskauskas,Mack,Kratzer,Salys,Kraskauskiene,Burns,Voelkel,Taraseviciene-Stewart:"Immunomodulatorystrategiespreventthedevelopmentofautoimmuneemphysema."in:Respiratoryresearch,Vol.11,pp.179,2010(PubMed).

新闻动态
行业前沿
技术文章
最新产品