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Agrisera/LEA45 | Late embryogenesis abundant protein 45 (affinity purified)/AS13 2758A/188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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Agrisera/LEA45 | Late embryogenesis abundant protein 45 (affinity purified)/AS13 2758A/
productinformation
Background

LEA(Lateembryogenesisabundant)proteinsareveryhydrophilicproteins,describedover25yearsagoasaccumulatingduringlatestagesofplantseeddevelopment.Foundinvegetativeplanttissuesfollowingexposuretoenvironmentalstress.Synonymes:PutativelateembryogenesisabundantproteinLEA.

Immunogen

recombinantLEA4-5fromArabidopsisthaliana:UniProt:Q9FG31,TAIR:AT5G06760, agroup4LEAprotein(Battaglia etal.,2008)

HostRabbit
ClonalityPolyclonal
Clone
PurityAffinitypurifiedseruminPBSpH7.4
FormatLyophilized
Quantity50µg
ReconstitutionForreconstitutionadd50µlofsterilewater.
Storage

storelyophilized/reconstitutedat-20°C;oncereconstitutedmakealiquotstoavoidrepeatedfreeze-thawcycles.Please,remembertospintubesbrieflypriortoopeningthemtoavoidanylossesthatmightoccurfromlyophilizedmaterialadheringtothecaporsidesofthetubes.

Testedapplicationswesternblot(WB)
Relatedproducts

AS132758|Anti-LEA4-5|Lateembryogenesisabundantprotein4-5,rabbitantibody
AS132756 |Anti-LEA6-3|Lateembryogenesisabundantprotein6-3,rabbitantibody

Plantproteinextractionbuffer

Secondaryantibodies

Additionalinformation
applicationinformation
Recommendeddilution

1:1000(WB)

Expected|apparentMW

16kDa

ConfirmedreactivityArabidopsisthaliana
PredictedreactivityArachishypogaea,Brassicasp.,Glycinemax,Medicagotruncatula,Phaseolusvulgaris,Thellungiellahalophila
Notreactivein

noconfirmedexceptionsfrompredictedreactivityarecurrentlyknown

Additionalinformation
Selectedreferences

tobeaddedwhenavailable,antibodyreleasedinAugust2016.


applicationinformation

western blot using affinity purified anti-AtLEA4-5 antibodies

ProteinextractswereobtainedfromArabidopsisthalianaseeds(extract),besiderecombinantAtLEA4-5loadedinindicatedamounts.Followingextractionbufferwasused:0.7Msucrose,0.5MTris-base,30mMHCl,50mMEDTA,0.1MKCl,2%β-mercaptoethanol,12mg/mlpoly-vinyl-poly-pyrrolidone(PVPP).Thisbufferwascomplementedwith2mLofequilbratedphenolbeforeextraction.Sampleswerecentrifugedandtheproteinphasewasrecovered;theextractedproteinswereprecipitatedwith0.1mammoniumacetatedissolvedinmethanol,centrifugedandthepelletwashedwithcold(-20°C)80%acetone.TheproteinpelletwasdissolvedinSDS-solubilizationbuffer(1%CHES,2%SDS,2%ß-mercaptoethanol,10%glycerol).ThirtyµgofseedproteinextractwasdenaturedwithLaemmlibufferat95°Cfor5minandproteinswereseparatedon15%SDS-PAGEandblotted1.5htonitrocellulosemembraneinaliquidtransfersystem.Blotswereblockedwith2%skimmilkONat4°Cwithagitation.AfterrinsingwithTBS,blotswereincubatedintheprimaryantibodyatadilutionof1:1000(anti-AtLEA4-5)for3hat4°Cwithagitation.Theantibodysolutionwasdecantedandtheblotwasrinsedbriefly,thenwashedoncefor15minand3timesfor5mininTBS-Tat4°Cwithagitation.Then,blotswereincubatedinsecondaryantibody(anti-rabbitIgGhorse-radishperoxidaseconjugate)dilutedto1:25000infor1hatRTwithagitation.Theblotwaswashedasaboveanddevelopedfor2minwithSuperSignalWestPicoChemiluminescentSubstrate,ThermoFisherScientific.Exposuretimewasfor30seconds.

CourtesyofDr.AlejandraCovarrubias,UNAM,MexicoCourtesyofDr.AlejandraA.Covarrubias,UNAM,Mexico


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