MouseIgG(H+L)Cross-AdsorbedSecondaryAntibody(A-11005)inIF
HumaniPSCStainingHumaniPSCswereculturedonglassslidesunderfeeder-freeconditionsinStemPro®hESCMedium(Product#A1000701).CellswerefixedandpermedwiththeImage-iT®Fixation/PermeABIlizationKit(Product#R37602).Oct4(green)expressionwasvisualizedusinganti-Oct4primaryAbandAlexaFluor®488secondaryAb(Product#A-11034).Tubulin(red)expressionwasvisualizedusinganti-tubulinprimaryAb(Product#32-2600)andAlexaFluor®594secondaryAb(Product#A-11005).Nuclei(blue)werelabeledwithNucBlue™FixedCellStain(Product#R37606).ImageswerecollectedontheFLoid™CellImagingStation(Product#4471136).
ProductDetails
| TESTEDAPPLICATIONS | DILUTION |
|---|---|
| FlowCytometry(Flow) | 1-10µg/mL |
| Immunocytochemistry(ICC) | 1-10µg/mL |
| Immunofluorescence(IF) | 1-10µg/mL |
| PUBLISHEDAPPLICATIONS | |
|---|---|
| Immunohistochemistry(Paraffin)(IHC(P)) | See6publicationsbelow |
| Immunocytochemistry(ICC) | See18publicationsbelow |
| Immunohistochemistry(Frozen)(IHC(F)) | See3publicationsbelow |
| MiscellaneousPubMed(MISC) | See190publicationsbelow |
| Immunohistochemistry(IHC) | See1publicationsbelow |
| Speciesreactivity | Mouse |
| Host/Isotype | Goat/IgG |
| Class | Polyclonal |
| Type | SecondaryAntibody |
| Immunogen | GammaImmunoglobinsHeavyandLightchains |
| Conjugate | AlexaFluor®594 |
| Excitation/EmissionProfile | Viewspectra  |
| Form | Liquid |
| Concentration | 2mg/ml |
| Purification | purified |
| Storagebuffer | PBS,pH7.5 |
| Contains | 5mMsodiumazide |
| Storageconditions | 4°C,storeindark |
| RRID | AB_2534073 |
| Target | IgG |
| CrossAdsorption | AgainsthumanIgGandhumanserumpriortoconjugation |
| AntibodyForm | WholeAntibody |
ProductSpecificInformation
Tominimizecross-reactivity,thesegoatanti-mouseIgGwholeantibodieshavebeencross-adsorbedagainsthumanIgGandhumanserumpriortoconjugation.Cross-adsorptionorpre-adsorptionisapurificationsteptoincreasespecificityoftheantibodyresultinginhighersensitivityandlessbackgroundstaining.Thesecondaryantibodysolutionispassedthroughacolumnmatrixcontainingimmobilizedserumproteinsfrompotentiallycross-reactivespecies.Onlythenonspecific-bindingsecondaryantibodiesarecapturedinthecolumn,andthehighlyspecificsecondariesflowthrough.Thebenefitsofthisextrastepareapparentinmultiplexing/multicolor-stainingexperiments(e.g.,flowcytometry)wherethereispotentialcross-reactivitywithotherprimaryantibodiesorintissue/cellfluorescentstainingexperimentswheretherearemaybethepresenceofendogenousimmunoglobulins.
AlexaFluordyesareamongthemosttrustedfluorescentdyesavailabletoday.Invitrogen™AlexaFluor594dyeisabright,red-fluorescentdyewithexcitationideallysuitedtothe594nmlaserline.Forstablesignalgenerationinimagingandflowcytometry,AlexaFluor594dyeispH-insensitiveoverawidemolarrange.Probeswithhighfluorescencequantumyieldandhighphotostabilityallowdetectionoflow-abundanceBIOLOGicalstructureswithgreatsensitivity.AlexaFluor594dyemoleculescanbeattachedtoproteinsathighmolarratioswithoutsignificantself-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.
