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蚂蚁淘在线 / 品牌中心 / BPS Bioscience / Bpsbioscience/Anti–5–hmC polyclonal antibody (rabbit)/25203/50 µg

Bpsbioscience/Anti–5–hmC polyclonal antibody (rabbit)/25203/50 µg

价格
￥7500.00
货号：25203
浏览量：127
品牌：BPS Bioscience
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商品描述
HostSpecies:Rabbit
Clonality:polyclonal
Background:
5–hydroxymethylcytosine(5–hmC)resultsfromtheenzymaticconversionof5–methylcytosineinto5–hydroxymethylcytosinebytheTETfamilyofiron–dependentoxygenases.5–hmCbaseswererecentlydiscoveredinmammalianDNA,inPurkinjeneurons,ingranulecellsandembryonicstemcellswheretheyarepresentathighlevels(upto0.6%oftotalnucleotidesinPurkinjecells).Recentreportsindicatethat5–hmCisabundantinbraintissue,especiallyinareasthatareassociatedwithhighercognitivefunctions.Preliminaryresultsindicatethat5–hmCmayhaveimportantrolesdistinctfrom5–mC.Althoughitspreciserolehasstilltobeshown,earlyevidencesuggests5–hydroxymethylcytosinemayrepresentanewanduniquepathwaytodemethylateDNAinvolvingarepairmechanismconverting5–hmCtocytosine.Duetothestructuralsimilaritybetween5–mCand5–hmC,thesebasesareexperimentallyalmostindistinguishable.Themostcommonlyusedmethodologies(e.g.enzymaticapproaches,bisulfitesequencing)donotdistinguish5mcfrom5–hmC.Thedevelopmentofspecificantibodiesappearstobethemostpowerfulwaytodistinguishandspecificallyenrichfor5–mCand5–hmCsequences.
Description:
Polyclonalantibodyraisedinrabbitagainst5–hydroxymethylcytosineconjugatedtoKLH.
AssayConditions:

hMeDIPresultsobtainedwiththeantibodydirectedagainst5-hmC
hMeDIP(hydroxymethylatedDNAIP)wasperformedusingtheantibodyagainst5-hydroxymethylcytosine(Cat.No.25203).DNAfrommouseEScellswaspreparedwiththeGenDNAmoduleofthehMeDIPkitandsonicatedtoobtainDNAfragmentsof300-500bp.OneμgofshearedDNAwasspikedwithunmethylated(C)methylated(mC),andhydroxymethylated(hmC)controls.hMeDIPwasperformedwith3.5μgoftherabbit5-hmCantibodyandtheimmunoprecipitatedDNAwasanalysedbyqPCRusingprimersspecificforthe3differentcontrolsequences.Figure1showsthattherabbitpolyclonalantibodyagainst5-hmCishighlyspecificforthe5-hmCbasemodification(noIPwithnon-methylatedormethylatedCcontrolfragments).

Determinationoftheantibodytiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainst5-hmC(cat.No.25203),crudeserumandflowthrough,inantigencoatedwells.Theantigenusedwasthe5-hmCbasecoupledtoBSA.Byplottingtheabsorbanceagainsttheantibodydilution,thetiteroftheantibodywasestimatedtobe1:2,800.

Dotblotanalysisusingtheantibodydirectedagainst5-hmC
Todemonstratethespecificityoftheantibodyagainst5-hmC(cat.No.25203),aDotblotanalysiswasperformedusinghmC,mCandCcontrols.Onehundredto4ng(equivalentof5to0.2pmolofC-bases)ofthecontrolswerespottedonamembrane(AmershamHybond-N+).Theantibodywasusedatadilutionof1:1,000.Figure3showsahighspecificityoftheantibodyforthehydroxymethylatedcontrol.

Concentration:
50µg
Formulation:
PBScontaining0.05%azideand0.05%ProClin300.
SpeciesReactivity:
Human,mouse,broadrange
Purification:
Affinitypurified
Immunogen:
nucleotide
Format:
Aqueousbuffersolution
Storage/StABIlity:

Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.

Application(s):
hMeDIP(3.5µg/IP)
ELISA(1:100–1:500)
DB(1:1000)
Notes:
Theoptimalantibodyconcentrationshouldbedeterminedbytheend–user.
Warning(s):
Avoidfreeze/thawcycles
ScientificCategory:
Antibodies
ProductType:
Antibody
Datashownislot-specific.Contactusforspecificinformationonotherlots.
BPS Bioscience生物素化，生物素标记通常用于蛋白质和其他目标分子的非放射性标记和纯化。生物素标记利用生物素（维生素H）与抗生物素蛋白或抗生蛋白链菌素之间异常强的相互作用。生物素与抗生物素蛋白或抗生蛋白链菌素的亲和力是已知的蛋白质和配体最强的非共价相互作用之一，在10 -15附近表现出解离常数（K d）。此外，这种相互作用可抵抗高或低pH，高盐浓度，极端温度和变性剂。生物素-亲和素相互作用的这些性质使其可用于纯化和/或检测目的蛋白质。BPS具有许多蛋白质，这些蛋白质已通过化学方法通过各种反应基团以及使用AviTag TM  技术进行酶促生物素标记。BPS的生物素化产品组合包括用于标记伯胺和仲胺，炔基，硫醇，羧基和磷酸基以及各种PEG基团的试剂。这种灵活性使研究人员能够满足他们所有的生物素化需求。