Rabbitpolyclonalantibody
PooledSerum- Size:
- 100µl
- Formulation:
- AffinityPurifiedfromPooledSerum
- Specificity:
- Rat,Mouse,Non-humanprimate
- Applications:
- WB1:1,000IF1:1,000IHC1:1,000
- Species:
- Rabbit
- GeneName:
- TH
- MolecularReference:
- ~60kDa
- CiteThisAntibody:
- PhosphoSolutionsCat#p1580-31,RRID:AB_2492278
Antigen/Purification:Collapse
Theantigenisaphosphopeptidecorrespondingtoaminoacidresiduessurroundingphospho-Ser31ofrattyrosinehydroxylase(TH).
Theantibodyispreparedfrompooledrabbitserumbyaffinitypurificationviasequentialchromatographyonphospho-anddephospho-peptideaffinitycolumns.
BIOLOGicalSignificance:Collapse
Tyrosinehydroxylase(TH)istherate-limitingenzymeinthesynthesisofthecatecholaminesdopamineandnorepinephrine.THantibodiescanthereforebeusedasMarkersfordopaminergicandnoradrenergicneuronsinavarietyofapplicationsincludingdepression,schizophrenia,Parkinson’sdiseaseanddrugabuse(Kishetal.,2001;Zhuetal.,2000;Zhuetal.,1999).THantibodiescanalsobeusedtoexplorebasicmechanismsofdopamineandnorepinephrinesignaling(Witkovskyetal.,2000;Salvatoreetal.,2001;Dunkleyetal.,2004).TheactivityofTHisalsoregulatedbyphosphorylation(Haycocketal.,1982;Haycocketal.,1992;Jedynaketal.,2002).Phospho-specificantibodiesforthephosphorylationsitesonTHcanbeusedtogreateffectinstudyingthisregulationandinidentifyingthecellsinwhichTHphosphorylationoccurs.
Storage
100µlin10mMHEPES(pH7.5),150mMNaCl,100µgpermlBSAand50%glycerol.Adequateamountofmaterialtoconduct10-miniWesternBlots.
Forlongtermstorage–20°Cisrecommended.Stableat–20°Cforatleast1year.
ProductSpecificReferences
Jorge-Finnigan,A.,Kleppe,R.,Jung-KC,K.,Ying,M.,Marie,M.,Rios-Mondragon,I.,Salvatore,M.F.,Saraste,J.andMartinez,A.,2017.Phosphorylationatserine31targetstyrosinehydroxylasetovesiclesfortransportalongmicrotubules.JournalofBiologicalChemistry,pp.jbc-M116.PMID:28637871
Bonito-Oliva,Alessandra,GiadaSpigolon,andGilbertoFisone.(2015)“RegulationofPre-andPostsynapticProteinPhosphorylationbyDopamineD2Receptors.”DopamineReceptorTechnologies.SpringerNewYork,2015.79-100.
Izumi,Y.,Ezumi,M.,Takada-Takatori,Y.,Akaike,A.,&Kume,T.(2014).Endogenousdopamineisinvolvedintheherbicideparaquat-induceddopaminergiccelldeath.ToxicologicalSciences,139(2),466-478.PMID:24743698
Calipari,E.S.,Ferris,M.J.,Melchior,J.R.,Bermejo,K.,Salahpour,A.,Roberts,D.,&Jones,S.R.(2014).Methylphenidateandcocaineself‐admiNISTrationproducedistinctdopamineterminalalterations.Addictionbiology,19(2),145-155.PMID:22458761
JedynakJP,AliSF,HaycockJW,HopeBT(2002)Acuteadministrationofcocaineregulatesthephosphorylationofserine-19,-31and-40intyrosinehydroxylase.JNeuRochem82:382-388.PMID:12124439
SalvatoreMF,WaymireJC,HaycockJW(2001)Depolarization-stimulatedcatecholaminebiosynthesis:involvementofproteinkinasesandtyrosinehydroxylasephosphorylationsitesinsitu.JNeurochem79:349-360.PMID:11677263
WitkovskyP,GabrielR,HaycockJW,MellerE(2000)Influenceoflightandneuralcircuitryontyrosinehydroxylasephosphorylationintheratretina.JChemNeuroanat19:105-116.PMID:10936746
PhosphoSolutions致力于重现性。寻找我们的“混合血清”图标。带有此图标的每种抗体均从其自身的血清库中纯化，以确保批次间的一致性。可重复性是科学研究中的一个大问题，这已经不是什么秘密了。针对特定蛋白质的抗体最初在实验中可能效果很好，但是当重新订购相同的抗体时，新小瓶中的“相同”产品会显示完全不同的结果。这种异常有多种可能的原因。购买抗体时要考虑的最重要方面之一是认识到相同的商业抗体通常可从许多不同的供应商处获得；确保您从同一家公司订购相同的产品以保持结果一致非常重要。但是，即使您这样做了，您仍可能会看到同一产品的批次间差异。发生这种情况是因为许多商业抗体公司从动物的不同血液甚至完全不同的动物中纯化不同批次。有了这样的可变性，难怪重现性是一个问题。下面显示了抗体信号强度和特异性如何随同一只兔子的出血而变化的示例：PhosphoSolutions相信有一个非常简单的方法可以解决可变性问题。PhosphoSolutions的解决方案是在抗体释放之前筛选和汇集从动物身上收集的所有血清。PhosphoSolutions在蛋白质印迹中筛选每只动物的每一次出血。如有必要，PhosphoSolutions然后从单个流血中纯化以测试亲和力和/或磷酸化特异性。一旦确定了含有高质量抗体的出血，PhosphoSolutions就会将它们合并成一个庞大的集合。因此，随后出售的所有抗体批次都是相同的，因为它们都是从相同的混合起始材料的相同等分试样中纯化而来的。通过这种做法，几乎消除了多克隆抗体的所有批次间差异。一旦抗体池耗尽，在抗体生产中是否需要格外小心？当然！这值得么？绝对地！PhosphoSolutions对有效的抗体充满热情，并认为您值得一次又一次依赖的研究工具。
