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Thermofisher/Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/A21202
  • Thermofisher/Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/A21202

Thermofisher/Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/A21202

價(jià)格: ￥2079.00 市場(chǎng)價(jià): 2495.00
貨號(hào): A21202
品牌: eBioscience
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    • MouseIgG(H+L)HighlyCross-AdsorbedSecondaryAntibody(A-21202)inIF

      ImmunofluorescenceanalysisofDonkeyanti-MouseIgG(H+L)SecondaryAntibody,AlexaFluor488conjugate(Product#A-21202)wasperformedusingMCF-7cellsstainedwithCytokeratin19MouseMonoclonalAntibody(Product#MA5-12613).Thecellswerefixedwith4%paraformaldehydefor10minutes,permeABIlizedwith0.1%Triton?X-100for10minutes,blockedwith1%BSAfor1hourandlabeledwithmouseprimaryantibody(1:250dilution)for3hoursatroomtemperature.Donkeyanti-MouseIgG(H+L)SecondaryAntibody,AlexaFluor488conjugatewasusedatconcentrationof0.2μg/mlinphosphatebufferedsalinecontaining0.2%BSAfor45minutesatroomtemperature,fordetectionofcytokeratin19inthemembrane(Panela:green).Nuclei(Panelb:blue)werestainedwithDAPIinSlowFade?GoldAntifadeMountant(Product#S36938).F-actinwasstainedwithRhodaminePhalloidin(Product#R415,1:300)(Panelc:red).Paneldrepresentsthecompositeimage.Nononspecificstainingwasobservedwiththesecondaryantibodyalone(panelf),orwithanisotypecontrol(panele).Theimageswerecapturedat60X 

      ProductDetails

      TESTEDAPPLICATIONSDILUTION
      Immunocytochemistry(ICC)0.2μg/ml
      Immunofluorescence(IF)1:2000
      Immunohistochemistry(IHC)1-10μg/ml
      PUBLISHEDAPPLICATIONS
      Immunohistochemistry(IHC)See6publicationsbelow
      Immunocytochemistry(ICC)See17publicationsbelow
      Immunohistochemistry(Paraffin)(IHC(P))See3publicationsbelow
      Immunohistochemistry(Frozen)(IHC(F))See5publicationsbelow
      FlowCytometry(Flow)See1publicationsbelow
      MiscellaneousPubMed(MISC)See36publicationsbelow
      SpeciesreactivityMouse
      Host/IsotypeDonkey/IgG
      ClassPolyclonal
      TypeSecondaryAntibody
      ImmunogenGammaImmunoglobinsHeavyandLightchains
      ConjugateAlexaFluor?488
      Excitation/EmissionProfileViewspectra 
      FormLiquid
      Concentration2mg/ml
      Purificationpurified
      StoragebufferPBS,pH7.5
      Contains5mMsodiumazide
      Storageconditions4°C,storeindark
      RRIDAB_141607
      TargetIgG
      AntibodyFormWholeAntibody

      ProductSpecificInformation

      Tominimizecross-reactivity,thesedonkeyanti-mouseIgGwholeantibodieshavebeenaffinity-purifiedandshowminimumcross-reactivitytobovine,chicken,goat,guineapig,hamster,horse,human,mouse,rat,andsheepserumproteins.Cross-adsorptionorpre-adsorptionisapurificationsteptoincreasespecificityoftheantibodyresultinginhighersensitivityandlessbackgroundstaining.Thesecondaryantibodysolutionispassedthroughacolumnmatrixcontainingimmobilizedserumproteinsfrompotentiallycross-reactivespecies.Onlythenonspecific-bindingsecondaryantibodiesarecapturedinthecolumn,andthehighlyspecificsecondariesflowthrough.Thebenefitsofthisextrastepareapparentinmultiplexing/multicolor-stainingexperiments(e.g.,flowcytometry)wherethereispotentialcross-reactivitywithotherprimaryantibodiesorintissue/cellfluorescentstainingexperimentswheretheremaybethepresenceofendogenousimmunoglobulins.

      AlexaFluordyesareamongthemosttrustedfluorescentdyesavailabletoday.Invitrogen?AlexaFluor488dyeisabright,green-fluorescentdyewithexcitationideallysuitedtothe488nmlaserline.Forstablesignalgenerationinimagingandflowcytometry,AlexaFluor488dyeispH-insensitiveoverawidemolarrange.Probeswithhighfluorescencequantumyieldandhighphotostabilityallowdetectionoflow-abundanceBIOLOGicalstructureswithgreatsensitivity.AlexaFluor488dyemoleculescanbeattachedtoproteinsathighmolarratioswithoutsignificantself-quenching,enablingbrighterconjugatesandmoresensitivedetection.Thedegreeoflabelingforeachconjugateistypically2-8fluorophoremoleculesperIgGmolecule;theexactdegreeoflabelingisindicatedonthecertificateofanalysisforeachproductlot.

      Usingconjugatesolutions:Centrifugetheproteinconjugatesolutionbrieflyinamicrocentrifugebeforeuse;addonlythesupernatanttotheexperiment.Thisstepwillhelpeliminateanyproteinaggregatesthatmayhaveformedduringstorage,therebyreducingnonspecificbackgroundstaining.Becausestainingprotocolsvarywithapplication,theappropriatedilutionofantibodyshouldbedeterminedempirically.Forthefluorophore-labeledantibodiesafinalconcentrationof1-10μg/mLshouldbesatisfactoryformostimmunohistochemistryandflowcytometryapplications.

      Background/TargetInformation

      WeofferanextensivelineofInvitrogen?secondaryantibodyconjugateswithwell-characterizedspecificityandlabeledwithawideselectionofpremiumfluorescentdyes,includingInvitrogen?AlexaFluor?fluorescentdyes.Fluorescentsecondaryantibodyconjugatesareusefulinthedetection,sorting,orpurificationofitsspecifiedtargetandidealforfluorescencemicroscopyandconfocallaserscanningmicroscopy,flowcytometry,andfluorescentwesterndetection.ThebreadthoffluorescentMarkersweofferallowsourreagentstobetailoredtoalmostanyfluorescentdetectionsystem.

      Secondaryantibodiesmaybeprovidedinthreeformats:wholeIgG,divalentF(ab')2fragments,andmonovalentFabfragments.Becauseofthehighdegreeofconservationinthestructureofmanyimmunoglobulindomains,mostclass-specificsecondaryantibodiesmustbeaffinity-purifiedandcross-adsorbedtoachieveminimalcross-reactionwithotherimmunoglobulins.

      Oursecondaryantibodyconjugatesaremostcommonlypreparedbyimmunizingthehostanimalwithapooledpopulationofimmunoglobulinsfromthetargetspeciesandcanbefurtherpurifiedandmodified(e.g.,immunoaffinitychromatography,antibodyfragmentation,labelconjugation,etc.)togeneratehighlyspecificreagents.Inthefirstroundofpurification,wholeimmunoglobulinsbindingtotheimmunizingantibodyarerecoveredandmainlyconsistofthe~150-kDaIgGclass.Furtherpurification,forexample,withProteinAorG,removesallunwantedimmunoglobulinclassesexcepttheaffinity-purifiedantibodiesthatreactwiththetarget-specificimmunoglobulinheavyand/orlightchains.

      ForResearchUseOnly.Notforuseindiagnosticprocedures.Notforresalewithoutexpressauthorization.
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