AAT Bioquest/Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction,AAT Bioquest,,AAT Bioquest,aatbio,AAT Bioquest抗体，试剂，血清，细胞 AAT Bioquest生物素,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：AAT Bioquest/Buccutite™ Rapid PE Antibody Labeling Kit *Microscale Optimized for Labeling 100 ug Antibody Per Reaction

产品编号：1310

市场价：¥143560.00

场地：美国(厂家直采)

产品分类：试剂盒>其它试剂盒>标记试剂盒>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$7178.00

品牌： AAT Bioquest

公司：AAT Bioquest

公司分类：

商品介绍

Overview

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Ex/Em(nm)	565/575
MW	N/A
CAS#	N/A
Solvent	DMSO
Storage	F/D/L
Category	ProteinBiochemistry Generalproteins
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R-Phycoerythrin(PE)isanorangefluorescentproteinwhichhasanexcitationwavelengthof565nmandanemissionwavelengthof575nm.AATBioquestoffersthisBuccutite™rapidlabelingkittofacilitatethePEconjugationstoantibodiesandotherproteinssuchasstreptavidinandothersecondaryreagents.Buccutite™PEConjugationKitprovidesarobustandconvenientmethodtoconjugateyourantibodieswithPE.ThekitincludesapreactivatedPEandreactionbuffer.TheconjugatedantibodycanbeusedinWB,ELISAandIHCapplications.Thiskitissufficientfor2labelingreactions,eachupto100ugofantibody.ConsideringthelargesizeofPE(240kDa),theamountofantibodyusedinalabelingreactionmustalwaysbelessthantheamountofPE.Thebestratioforanynewantibodyreagentmustbedeterminedbyexperimentationbut50-60ugofIgGantibodyforevery100ugofPEusuallygivesoptimalresults.OurkitprovidespreactivatedPEtofacilitatethePEconjugationstoantibodiesandotherproteinssuchasstreptavidinandothersecondaryreagents.OurpreactivatedPEisreadytoconjugate,givingmuchhigheryieldthantheconventionallytediousSMCC-basedconjugationchemistry.Inaddition,ourpreactivatedPEisconjugatedtoaproteinviaitsaminogroupthatisabundantinproteinswhileSMCCchemistrytargetsthethiolgroupthathastoberegeneratedbythereductionofantibodies.

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

Prepareantibodysolution:
Forlabeling100μgantibody(assumingthetargetantibodyconcentrationis1mg/mL),mix5μL(5%ofthetotalreactionvolume)ofReactionBuffer(ComponentC)with100μLofthetargetantibodysolution.
Note1.Ifyouhaveadifferentantibodyconcentration,adjusttheantibodyvolumeaccordinglytomake~100µgantibodyavailableforyourlabelingreaction.
Note2:Theantibodyshouldbedissolvedin1Xphosphatebufferedsaline(PBS),pH7.2-7.4;Iftheantibodyisdissolvedinglycinebuffer,itmustbedialyzedagainst1XPBS,pH7.2-7.4,oruseAmiconUltra-0.5,Ultracel-10Membrane,10kDa(cat#UFC501008fromMillipore)toremovefreeaminesorammoniumsalts(suchasammoniumsulfateandammoniumacetate)thatarewidelyusedforproteinprecipitation.
Note3:ImpureantibodiesorantibodiesstABIlizedwithbovineserumalbumin(BSA)orgelatinwillnotbelabeledwell.
Note4:TheAntibody–Buccutite™MTAreactionefficiencyissignificantlyreducediftheantibodyconcentrationislessthan1mg/mL.Foroptimallabelingefficiencythefinalantibodyconcentrationrangeof1-10mg/mLisrecommended.

RunAntibody-Buccutite™MTAreaction:
1. AddtheantibodysolutiondirectlyintothevialofBuccutite™MTA(ComponentB),andmixthemwellbyrepeatedlypipettingforafewtimesorvortexthevialforafewseconds.
2. KeeptheAntibody-Buccutite™MTAreactionmixtureatroomtemperaturefor30-60minutes.
  Note:TheAntibody-Buccutite™MTAreactionmixturecanberotatedorshakenforlongertimeifdesired.

PreparespincolumnforAntibody-Buccutite™MTApurification:
1. Inverttheprovidedspincolumn(ComponentD)severaltimestore-sUSPendthesettledgelandremoveanybubbles.
2. Snapoffthetipandplacethecolumninawashingtube(2mL,notprovided).Removethecaptoallowtheexcesspackingbuffertodrainbygravitytothetopofthegelbed.Ifcolumndoesnotbegintoflow,pushcapbackintocolumnandremoveitagaintostarttheflow.Discardthedrainedbuffer,andthenplacethecolumnbackintotheWashingTube.However,centrifugeimmediatelyifthecolumnisplacedintoa12x75mmtesttube(notprovided).
3. Centrifugefor2minutesinaswingingbucketcentrifugeat1,000xg(seeCentrifugationNotessection)toremovethepackingbuffer.Discardthebuffer.
4. Apply1-2mL1XPBS(pH7.2-7.4)tothecolumn.AftereachapplicationofPBS,letthebufferdrainoutbygravity,orcentrifugethecolumnfor2minutestoremovethebuffer.Discardthebufferfromthecollectiontube.Repeatthisprocessfor3-4times.
5. Centrifugefor2minutesinaswingingbucketcentrifugeat1,000xg(seeCentrifugationNotessection)toremovethepackingbuffer.Discardthebuffer.

PurifytheAb-Buccutite™MTAsolution:
1. Placethecolumn(fromStep3.5)inacleanCollectingTube(1.5mL,notprovided).Carefullyloadthesample(~105μL,fromStep2.2)directlytothecenterofthecolumn.
2. AfterloADIngthesample,add5μLof1XPBS(pH7.2-7.4)tomakethetotalvolumeof110μL.Centrifugethecolumnfor5-6minutesat1,000xg,andcollectthesolutionthatcontainsthedesiredprotein-Buccutite™MTAsolution.

MakeAb-PEorPETandemconjugation:
1. MixwholevialofBuccutite™FOL-ActivatedPEorPETandem(ComponentA)withthepurifiedAb-Buccutite™MTAsolution(fromStep4.2),androtatethemixturefor1houratroomtemperature.
2. TheAb-PEorPETandemconjugateisnowreadytouse.
  Note1:Forimmediateuse,theAb-PEorPETandemconjugateneedbedilutedwiththebufferofyourchoice.
  Note2:Theconcentrationoftheconjugateisabout0.5~0.6mgAb/mLifstartwith100uL1mg/mlantibodysolution.

References&Citations

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1. ZhaoKH,SuP,LiJ,TuJM,ZhouM,BubenzerC,ScheerH.(2006)Chromophoreattachmenttophycobiliproteinbeta-subunits:phycocyanobilin:cysteine-beta84phycobiliproteinlyaseactivityofCpeS-likeproteinfromAnabaenaSp.PCC7120.JBiolChem,281,8573.

2. PetrasekZ,SchmittFJ,TheissC,HuyerJ,ChenM,LarkumA,EichlerHJ,KemnitzK,EckertHJ.(2005)ExcitationenergytransferfromphycobiliproteintochlorophylldinintactcellsofAcaryochlorismarinastudiedbytime-andwavelength-resolvedfluorescencespectroscopy.PhotochemPhotobiolSci,4,1016.

3. LoosD,CotletM,DeSchryverF,HabuchiS,HofkensJ.(2004)Single-moleculespectroscopyselectivelyprobesdonorandacceptorchromophoresinthephycobiliproteinallophycocyanin.BiophysJ,87,2598.

4. PrasannaR,PrasannaBM,MohammadiSA,SinghPK.(2003)EvaluationofTolypothrixgermplasmforphycobiliproteincontent.FoliaMicrobiol(Praha),48,59.

5. PrasannaR,DharDW,DominicTK,TiwariON,SinghPK.(2003)IsolationandcharacterisationofphycobiliproteinrichmutantofcyanobacteriumSynechocystissp.ActaBiolHung,54,113.

6. WuP.(2000)[Phycobiliproteinandfluorescenceimmunologicalassay].ShengLiKeXueJinZhan,31,82.

7. NoubirS,LuqueI,OchoadeAldaJA,PerewoskaI,TandeaudeMarsacN,CobleyJG,HoumardJ.(2002)Co-ordinatedexpressionofphycobiliproteinoperonsinthechromaticallyadaptingcyanobacteriumCalothrixPCC7601:aroleforRcaDandRcaG.MolMicrobiol,43,749.

8. TingCS,RocapG,KingJ,ChisholmSW.(2001)PhycobiliproteingenesofthemarinephotosyntheticprokaryoteProchlorococcus:evidenceforrapidevolutionofgeneticheterogeneity.MicroBIOLOGy,147,3171.

9. TriantafilouK,TriantafilouM,WilsonKM.(2000)Phycobiliprotein-Fabconjugatesasprobesforsingleparticlefluorescenceimaging.Cytometry,41,226.

10. ZhaoKH,DengMG,ZhengM,ZhouM,ParbelA,StorfM,MeyerM,StrohmannB,ScheerH.(2000)Novelactivityofaphycobiliproteinlyase:boththeattachmentofphycocyanobilinandtheisomerizationtophycoviolobilinarecatalyzedbytheproteinsPecEandPecFencodedbythephycoerythrocyaninoperon.FEBSLett,469,9.

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品牌介绍

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