Agrisera/ZEP | Zeaxanthin Epoxidase/AS08 289/


productinformation
Background		ZeaxanthinEpoxidase(ZEP)catalysestheO2-/NAPDHdependentepoxidationofzeaxanthintoviolaxanthinviaantheraxanthinatthestromalsideofthethylakoidmembraneofgreenplants.ZEPalsofunctionsinthefirststepofthebiosynthesisoftheABIoticstresshormoneabscisicacid(ABA).Synonymes:ProteinABADeficient1,AtABA1,ProteinImpairedinBABA-InducedSterility3,ProteinLowExpressionofOsmoticStress-ResponsiveGenes6,ProteinNon-PhotochemicalQuenching2.
Immunogen		KLH-conjugatedsyntheticpeptidederivedfromZEPsequenceofArabidopsisthalianaQ9FGC7 AT5G67030
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		50µl
Reconstitution		Forreconstitutionadd50µlofsterilewater.
Storage		storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.
Testedapplications		westernblot(WB)
Relatedproducts
Additionalinformation

applicationinformation
Recommendeddilution		1:1000-1:5000withstandardECL(WB)
Expected\|apparentMW		73.8\|70-75kDa(Arabidopsisthaliana)
Confirmedreactivity		Arabidopsisthaliana
Predictedreactivity		dicotsincluding:Brassicarapa,Vitisvinifera
Notreactivein		Glycinemax,Hordeumvulgare,Spinaciaoleracea
Additionalinformation
Selectedreferences		tobeaddedwhenavailable

applicationexample

western blot using anti-ZEP antibodies

10µgoftotalproteinfromsamplessuchas Arabidopsisthaliana leaf (1) ,Hordeumvulgare leaf (2)wereextractedwithProteinExtractionBufferPEB(AS08300).Samplesweredilutedwith1Xsamplebuffer(NuPAGELDSsamplebuffer(Invitrogen)supplementedwith50mMDTTandheatat70°Cfor5minandkeeptonicebeforeloADIng.Proteinsampleswereseparatedon4-12% BoltPlusgels, LDS-PAGEandblottedfor 70minutes toPVDFusingtanktransfer.Blotswereblockedimmediatelyfollowingtransferin2%blockingreagent(GERPN2125;Healthcare)or5%non-fatmilkdissolvedin20mMTris,137mMsodiumchloridepH7.6with0.1%(v/v)Tween-20(TBS-T)for1hatroomtemperaturewithagitation.Blotswereincubatedintheprimaryantibodyatadilutionof1:10000(inblockingreagent)for1hatroomtemperaturewithagitation.Theantibodysolutionwasdecantedandtheblotwasrinsedbrieflytwice,andthenwashed1x15minand3x5minwithTBS-Tatroomtemperaturewithagitation.Blotswereincubatedinsecondaryantibody(anti-rabbitIgGhorseradishperoxidaseconjugated,recommendedsecondaryantibody AS101489,Agrisera)dilutedto1:25000inblockingreagentfor1hatroomtemperaturewithagitation.Theblotswerewashedasabove.Theblotwasdevelopedfor5minwithTMA-6(lumigen)detectionreagentaccordingthemanufacturersinstructions.ImagesoftheblotswereobtainedusingaCCDimager(VersaDocMP4000) andQuantityOnesoftware(Bio-Rad).Exposuretimewas3minutes.

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Agrisera/ZEP | Zeaxanthin Epoxidase/AS08 289/