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Thermofisher/Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/A21202188bio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
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Thermofisher/Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488/A21202

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
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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