Agrisera/Clathrin heavychain 1,2/AS10 690/


productinformation
Background		Clathrinisaproteininvolvedinintracellulartraffickingandplaysamajorroleintheformationofcoatedvesicles.Itconsistsofthreeclathrinheavychainsandthreelightchains.Clathrin-coatedvesicles(CCV)selectivelysortcargoatthecellmembrane,trans-Golginetwork,andendosomalcompartmentsformultiplemembranetrafficpathways.
Immunogen		KLH-conjugatedpeptidederivedfromavailableplantclathrinheavychainsequencesincludingArabidopsisthalianaclathrinheavychain1UniProt:Q0WNJ6,TAIR:At3g11130,clathrinheavychain2UniProt:Q0WLB5,TAIR:At3g08530
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Affinitypurifiedserum
Format		Lyophilized
Quantity		100µg
Reconstitution		Forreconstitutionadd100µlofsterilewater
Storage		storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.
Testedapplications		westernblot(WB),immunolocalization(IL),immunofluorescence(IF)
Relatedproducts		AS10680\|Tubulinalphachain AS10681\|Tubulinbetachain Plantandalgalproteinextractionbuffer Secondaryantibodies
Additionalinformation

applicationinformation
Recommendeddilution		1:2000withstandardECL(WB),1:2400(IL),1:400(IF)
Expected\|apparentMW		193\|170kDa(Arabidopsisthaliana)
Confirmedreactivity		Arabidopsisthaliana,Chlamydomonasreinhardtii,Nicotianatabacum
Predictedreactivity		Amborellatrichopoda,Brassicanapus,Capsellarubella,Citrusaurantiumvar.sinensis,Eucalyptusgrandis, Glycinemax,Chlorellavariabilis, Leucaenaglauca,Lotusjaponicus, Medicagotribuloides, Mimulusguttatus,Musamalaccensis, Oryzasativa,Panicumitalicum, Physcomitrellapatens,Phaseolusvulgaris,Pisumsativum,Populusbalsamifera,Ricinuscommunis,Selaginellamoellendorffii,Sisymbriumsalsugineum,Solanumlycopersicum,Theobromacacao,Triticumaestivum,Vitisvinifera
Notreactivein		Nicotianabenthamiana
Additionalinformation		Please,notethatfreshsampleswillprovidebetterrestuls(seeimagebelow).
Selectedreferences		Wattelet-Boyeretal.(2016).EnrichmentofhydroxylatedC24-andC26-acyl-chainsphingolipidsmediatesPIN2apicalsortingattrans-Golginetworksubdomains.NatCommun.2016Sep29;7:12788.doi:10.1038/ncomms12788. Derbyshireetal.(2015).ProteomicAnalysisofMicrotubuleInteractingProteinsovertheCourseofXylemTrachearyElementFormationinArabidopsis.PlantCell.2015Oct2.pii:tpc.15.00314. Gronesetal.(2015).Auxin-bindingpocketofABP1iscrucialforitsgain-of-functioncellularanddevelopmentalroles.JExpBot.2015Apr28.pii:erv177. McLoughlinetal.(2013).IdentificationofnovelcandidatephosphatidicacidbindingproteinsinvolvedinthesaltstressresponseofArabidopsisthalianaroots.BiochemJ.Jan17.

applicationexample

western blot using anti-plant clathrin heavy chain antibody

10or20µloffreshlypreparedtotalproteinfromArabidopsisthalianasuspensionculture(A),10or20µloftotalprotein(olderextract)fromArabidopsisthalianasuspensionculture(B),10or20µloftotalleafextractfromNicotianatabacum(C),10or20µloffreshlypreparedtotalproteinfromNicotianatabacumleafprotoplasts(D),10or20µltotalproteinfromolderextractofNicotianatabacumleafprotoplasts(E),extractedwithbuffercontaining100mMTris(pH7.8),200mMNaCl,1mMEDTA,2%(v/v)beta-mercaptoethanoland0.2%(v/v)TritonX-100,wereseparatedon8%SDS-PAGEandblotted2htonitrocellulose(semidryblotat200mAandRT).Blotswereblockedwith5%(w/v)milkpowderand1%(w/v)BSAovernightat4°Cwithagitation.Blotwasincubatedintheprimaryantibodyatadilutionof1:2000for1hatRTwithagitation.Theantibodysolutionwasdecantedandtheblotwasrinsedbrieflytwice,thenwashedoncefor15minand3timesfor5mininTBS-TatRTwithagitation.Blotwasincubatedinsecondaryantibody(anti-rabbitIgGHRPconjugated)dilutedto1:10000inblockingsolutionfor45min.atRTwithagitation.Theblotwaswashedasaboveanddevelopedfor5minwithECLaccordingtothemanufacturersinstructions.Exposuretimewas120seconds.

10orCourtesyFabianKünzl,UniversityofTuebingen,Germany

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Agrisera/Clathrin heavychain 1,2/AS10 690/