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GenWay/Mouse IgG Antibody (HRP) (rat adsorbed)/GWB-545470/0.5 mg
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4000-520-616
Description:
ShortDescription:GOATANTIMOUSEIgGHRP(RATADSORBED)AdditionalInformation:
| Name | MouseIgGAntibody(HRP)(ratadsorbed) |
|---|---|
| RelatedProductNames | GOATANTIMOUSEIgG:HRP(RATADSORBED)IgGIGHG1 |
| GeneSymbol | Ighg1 |
| GeneFamily | IGH |
| Geneid | 16017 |
| AliasSymbols | IgG1,Igh-4,VH7183,Ighg1 |
| ProteinName | Iggamma-1chainCregion,membrane-boundform |
| DescriptionofTarget | GOATANTIMOUSEIgG:HRP(RATADSORBED) |
| SwissprotID | P01869 |
| SpeciesReactivity | Mouse |
| Host | Goat |
| Isotype | PolyclonalIgG |
| Conjugation | HRP |
| Replacement | Thisantibodymayreplaceitemsc-52003fromSantaCruzBiotechnology. |
| Immunogen | MouseIgG |
| Homology | Mouse |
| ProductFormat | PurifiedIgGconjugatedtoHorserADIshPeroxidase(HRP)-liquid |
| SwissProtNumber | P01869 |
| Specificity | IgG |
| Concentration | 0.5mg/ml |
| Applications | C,E,P |
| ApplicationInfo | Immunohistology-Frozen: 1/50 ELISA: 1/500-1/1000 |
| Clonality | Polyclonal |
| Format | Solution:Phosphatebufferedsaline,Stabalizer:0.01%Thiomersal,HRPStABIliser,Form:PurifiedIgGconjugatedtoHorseradishPeroxidase(HRP)-liquid |
| Stability | 18monthsfromdateofdespatch. |
| ReconstitutionandStorage | Storeat+4oC.DONOTFREEZE. Thisproductshouldbestoredundiluted.Shouldthisproductcontainaprecipitatewerecommendmicrocentrifugationbeforeuse. |
| Reactivity | Mouse |
| Storage | Storeat+4°C.DONOTFREEZE.Thisproductshouldbestoredundiluted.Shouldthisproductcontainaprecipitatewerecommendmicrocentrifugationbeforeuse. |
| ProtocolInformation | Citation:1:DennisA,KudoT,KruidenierL,GirardF,CrepinVF,MacDonaldTT,FrankelG,WilesS.Thep50subunitofNF-kappaBiscriticalforinvivoclearanceofthenoninvasiveentericpathogenCitrobacterrodentium.InfectImmun.2008Nov;76(11):4978-88.Epub2008Aug11.PubMedPMID:18694964;PubMedCentralPMCID:PMC2573329. Species:ThebacterialstrainCitrobacterrodentium,Mouse ExperimentName:1.Immunohistochemistryand2.DeterminationofC.rodentium-specificantibodyresponses. ExperimentBackground:1.Inthiswork,Alisonetal.characterizedtheimmuneresponseandpathologyofmicelackingthep50subunitofthetranscriptionfactornuclearfactorkappaB(NF-ĸB)duringC.rodentiuminfection.Theyshowedthatp50–/–miceareunabletoclearC.rodentiuminfection.2.Micelackingp50showdelayedandreducedimmunecellinfiltrationintocolonictissueduringC.rodentiuminfection.ImmunecellinfiltratesweremeasuredduringC.rodentiuminfectionofp50+/+miceandp50–/–miceusingfrozensectionsofmurinecolonincubatedwithantibodiesagainstCD3,CD4,CD8,CD11casaMarkerfordendriticcells[DC],andF4/80asamarkerformacrophages[Mø] ExperimentalSteps:1Immunohistochemistry:CryostatsectionsofmurinecolonwererehydratedinTris-bufferedsaline(TBS)for5minandthenincubatedwithantibodiesagainstCD3,CD4,CD8,F4/80,CD11c,NK1.1,orCD45R(Oxford,UnitedKingdom)ata1:10to1:100dilutionfor1h.ThesectionsweregentlywashedthreetimeswithTBSbeforetheadditionofbiotinylatedanti-ratoranti-hamsterimmunoglobulinG(IgG),asappropriate,atadilutionof1:200with4%(vol/vol)normalmurineserumforblocking(SeraLaboratoriesInternational,HorstedKeynes,UnitedKingdom)for30min.Afterthewashing,a1:200dilutionof0.1%avidin-peroxidase(Sigma-Aldrich)wasaddedfor30minbeforefurtherwashingandtheadditionofdiaminobenzadinesubstrate(Sigma-Aldrich)for5to10min.ThereactionwasstoppedwithexcessTBS,andthesectionswerecounterstainedwithhematoxylinandmounted.Acontrolslideusingnoprimaryantibodywasalsousedtoshowendogenousperoxidase-containingcells.Stainedcellpopulationswerecountedinfiverandomlyselectedfieldspersection,andthedatawereexpressedasthenumberofTcellsper250µm2oflaminapropria.2DeterminationofC.rodentium-specificantibodyresponses.An18-hcultureofC.rodentiumwasresUSPendedinPBSplus1%bacterialproteaseinhibitor(Sigma-AldrichLtd.,Dorset,UnitedKingdom)toanopticaldensityat600nm(OD600)of1.0andthenheatkilledat60°Cfor1h.A1:50PBSdilutionofthisculturewasaddedtoELISAplates(BDBiosciences,Oxford,UnitedKingdom)at100µlperwellbeforeincubationat4°Covernight.Theplateswerewashedthreetimeswith200µlperwellPBSplus0.05%Tweenandthenblockedfor1hwith200µlperwell1%bovineserumalbumininPBSblockingbuffer.Atselectedtimepointspostinoculation,bloodwascollectedfromthemicebycardiacpunctureandcentrifugedat8,000xgtoseparateredbloodcellsfromserum.TheC.rodentium-coatedELISAplateswerewashed.andthenserumsamplesdiluted1:100withPBSwereaddedinduplicatewells(50µlperwell)andincubatedatroomtemperaturefor2h.Theplateswerewashedasbeforeandthenincubatedatroomtemperaturefor2hwith100µlperwell1:2,000blockingbufferdilutionsofratanti-mouseIgG,IgG2b,IgG3,orIgMconjugatedtoalkalinephosphatase(AbD,Oxford,UnitedKingdom).Aftertheplateswerewashed,antibodiesweredetectedwith100µlperwell1-mg/mlp-nitrophenylphosphate(Sigma-AldrichLtd.,Dorset,UnitedKingdom)for5to10minbeforethereactionwasstoppedwith25µlperwell3NNaOH.AbsorbancewasdeterminedusingaVersaMaxmicroplatereaderwithSoftmaxPROsoftware(MolecularDevices,California)at405nm. OtherReagentsUsed:kanamycin,nalidixicacid NumberOfProtocols:2 |
| LeadTime | Domestic:within2-3weeksdelivery International:2-3weeks |
| IntendedUse | ResearchUseOnly |
| :: | PreservativeStabilisers:0.01%Thiomersal 1%BovineSerumAlbumin HRPStabiliser(BUF052A) AntiserumPreparation:AntiseratomouseIgGwereraisedbyrepeatedimmunisationofgoatswithhighlypurifiedantigen.PurifiedIgGwaspreparedbyaffinitychromatography. |
| :: | ApproxProteinConc:IgGconcentration0.5mg/ml BufferSolutions:Phosphatebufferedsaline |
