RabbitIgG(H+L)Cross-AdsorbedSecondaryAntibody(A-11008)inIF
ImmunofluorescenceanalysisofGoatanti-RabbitIgG(H+L)Cross-AdsorbedSecondaryAntibodyAlexaFluor®488conjugatewasperformedusingHeLacellsstainedwithalphaTubulinRabbitPolyclonalAntibody(Product#PA5-16891).Thecellswerefixedwith4%paraformaldehydefor10minutes,permeABIlizedwith0.1%Triton™X-100for10minutes,blockedwith1%BSAfor1hourandlabeledwith2µg/mlRabbitprimaryantibodyfor3hoursatroomtemperature.Goatanti-RabbitIgG(H+L)Cross-AdsorbedSecondaryAntibodyAlexaFluor®488conjugate(Product#A-11008)wasusedataconcentrationof4µg/mlinphosphatebufferedsalinecontaining0.2%BSAfor45minutesatroomtemperature,fordetectionofalphaTubulininthecytoplasm(Panela:green).Nuclei(Panelb:blue)werestainedwithDAPIinSlowFade®GoldAntifadeMountant(Product#S36938).F-actinwasstainedwithRhodaminePhalloidin(Product#R415,1:300)(Panelc:red).Paneldrepresentsthecompositeimage.Nononspecificstainingwasobservedwiththesecondaryantibodyalone(panelf),orwithanisotypecontrol(panele).Theimageswerecapturedat60Xmagnification.
ProductDetails
| TESTEDAPPLICATIONS | DILUTION |
|---|---|
| FlowCytometry(Flow) | 1-10µg/mL |
| Immunocytochemistry(ICC) | 4µg/mL |
| Immunofluorescence(IF) | 4µg/mL |
| PUBLISHEDAPPLICATIONS | |
|---|---|
| Immunohistochemistry(Frozen)(IHC(F)) | See20publicationsbelow |
| Immunohistochemistry(IHC) | See20publicationsbelow |
| Immunocytochemistry(ICC) | See26publicationsbelow |
| Immunohistochemistry(Paraffin)(IHC(P)) | See8publicationsbelow |
| FlowCytometry(Flow) | See2publicationsbelow |
| WesternBlot(WB) | See3publicationsbelow |
| Immunohistochemistry-FreeFloating(IHC(Free)) | See1publicationsbelow |
| ChIPassay(ChIP) | See1publicationsbelow |
| MiscellaneousPubMed(MISC) | See472publicationsbelow |
| Speciesreactivity | Rabbit |
| Host/Isotype | Goat/IgG |
| Class | Polyclonal |
| Type | SecondaryAntibody |
| Immunogen | GammaImmunoglobinsHeavyandLightchains |
| Conjugate | AlexaFluor®488 |
| Excitation/EmissionProfile | Viewspectra  |
| Form | liquid |
| Concentration | 2mg/ml |
| Purification | purified |
| Storagebuffer | PBS,pH7.5 |
| Contains | 5mMsodiumazide |
| Storageconditions | 4°C,storeindark |
| RRID | AB_143165 |
| Target | IgG |
| CrossAdsorption | AgainsthumanIgG,humanserum,mouseIgG,mouseserumandbovineserum |
| AntibodyForm | WholeAntibody |
ProductSpecificInformation
AlexaFluordyesareamongthemosttrustedfluorescentdyesavailabletoday.Invitrogen™AlexaFluor™488dyeisabright,green-fluorescentdyewithexcitationideallysuitedtothe488nmlaserline.Forstablesignalgenerationinimagingandflowcytometry,AlexaFluor488dyeispH-insensitiveoverawidemolarrange.Probeswithhighfluorescencequantumyieldandhighphotostabilityallowdetectionoflow-abundanceBIOLOGicalstructureswithgreatsensitivity.AlexaFluor488dyemoleculescanbeattachedtoproteinsathighmolarratioswithoutsignificantself-quenching,enablingbrighterconjugatesandmoresensitivedetection.Thedegreeoflabelingforeachconjugateistypically2-8fluorophoremoleculesperIgGmolecule;theexactdegreeoflabelingisindicatedonthecertificateofanalysisforeachproductlot.
Thegoatanti-rabbitIgGwholeantibodyconjugatesaremostcommonlypreparedbyimmunizingthehostanimalwithapooledpopulationofimmunoglobulinsfromthetargetspeciesandcanbefurtherpurifiedandmodified(e.g.,immunoaffinitychromatography,antibodyfragmentation,labelconjugation,etc.)togeneratehighlyspecificreagents.Inthefirstroundofpurification,wholeimmunoglobulinsbindingtotheimmunizingantibodyarerecoveredandmainlyconsistofthe~150-kDaIgGclass.FurtherpurificationwithProteinAorGremovesallimmunoglobulinclassesexceptIgGsuchthattheaffinity-purifiedantibodiesreactwithIgGheavychainsandallclassesofimmunoglobulinlightchainsfromrabbit.Tominimizecross-reactivity,thesegoatanti-rabbitwholeantibodieshavebeencross-adsorbedagainsthumanIgG,humanserum,mouseIgG,mouseserum,andbovineserum.Cross-adsorptionorpre-adsorptionisapurificationsteptoincreasespecificityoftheantibodyresultinginhighersensitivityandlessbackgroundstaining.Thesecondaryantibodysolutionispassedthroughacolumnmatrixcontainingimmobilizedserumproteinsfrompotentiallycross-reactivespecies.Onlythenonspecific-bindingsecondaryantibodiesarecapturedinthecolumn,andthehighlyspecificsecondariesflowthrough.Thebenefitsofthisextrastepareapparentinmultiplexing/multicolor-stainingexperimentswherethereispotentialcross-reactivitywithotherprimaryantibodiesorinimmunohistochemistryexperimentswheretherearemaybethepresenceofendogenousimmunoglobulins.Forahighlycross-adsorbedsecondaryantibodyequivalent(orequivalentsecondaryantibodypreparation),pleaseseeproductcatalognumber:A11034.
Usingconjugatesolutions:Centrifugetheproteinconjugatesolutionbrieflyinamicrocentrifugebeforeuse;addonlythesupernatanttotheexperiment.Thisstepwilleliminateanyproteinaggregatesthatmayhaveformedduringstorage,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.
ForResearchUseOnly.Notforuseindiagnosticprocedures.Notforresalewithoutexpressauthorization.
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（1）优级纯试剂 亦称保证试剂，为一级品，纯度高，杂质极少，主要用于精密分析和科学研究，常以GR表示。
（2）分析纯试剂 亦称分析试剂，为二级品，纯度略低于优级纯，杂质含量略高于优级纯，适用于重要分析和一般性研究工作，常以AR表示。
（3）化学纯试剂 为三级品，纯度较分析纯差，但高于实验试剂，适用于工厂、学校一般性的分析工作，常以CP表示。
（4）实验试剂 为四级品，纯度比化学纯差，但比工业品纯度高，主要用于一般化学实验，不能用于分析工作，常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定，化学试剂的不同等级分别用各种不同的颜色来标志，见表1。
表1 我国化学试剂的等级及标志
bhclna2so4nano3na2co3
chclnaohna2co3nacl
dba(oh)2nahco3alcl3nahso4
刚入这行，谢谢大家
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3
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