Bpsbioscience/Anti–5–hmC polyclonal antibody (rabbit)/25205/100 µl188bio精品生物—专注于实验室精品爆款的电商平台

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–hydroxymethylcytosine(5–hmC)conjugatedtoKLH

AssayConditions:

Determinationofthe5-hmCrabbitpolyclonalantibodytiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftherabbitpolyclonalantibodydirectedagainst5-hmCinantigencoatedwells.TheantigenusedwasBSAcoupledtothe5-hmCbase.Byplottingtheabsorbanceagainsttheantibodydilution,thetiteroftheantibodywasestimatedtobe1:3,500.

AnhydroxymethylatedDNAIP(hMeDIP)wasperformedusingtherabbitpolyclonalantibodydirectedagainst5-hydroxymethylcytosine(Cat.No.25205).
TheIgGisotypeantibodiesfromrabbitwereusedasnegativecontrol.TheDNAwaspreparedwiththeGenDNAmoduleofthehMeDIPkitandsonicatedwtohaveDNAfragmentsof300-500bp.1μgofhumanHeLacellsDNAwerespikedwithnon-methylated,methylated,andhydroxymethylatedfragments.TheimmunoprecipitatedmaterialhasbeenanalysedbyqPCRusingtheprimerpairspecificforthe3differentcontrolsequences.Theobtainedresultsshowthattherabbitpolyclonalfor5-hmCishighlyspecificforthisbasemodification(noIPwithnon-methylatedormethylatedCbasescontainingfragments).

Dotblotanalysisofthe5-hmCrabbitpolyclonalantibodywiththeC,mCandhmCPCRcontrols
100to4ng(equivalentof5to0.2pmolofC-bases)ofthehmC,mCandCPCRcontrolswerespottedonamembrane(AmershamHybond-N+).Themembranewasincubatedwiththerabbit5-hydroxymethylcytosinepolyclonalantibody(dilution1:200).Themembraneswereexposedfor30seconds.

Concentration:

100µl

Formulation:

Wholeantiserumfromrabbitcontaining0.05%azide

SpeciesReactivity:

Human,mouse,broadrange

Purification:

Wholeserum

Immunogen:

nucleotide

Format:

aqueoussolution

Storage/Stability:

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

Application(s):

hMeDIP(2.5µg/IP)
ELISA(1:500)
DB(1:200)

Notes:

Theoptimalantibodyconcentrationshouldbedeterminedbytheend–user.

Warning(s):

Avoidfreeze/thawcycles

ScientificCategory:

Antibodies

ProductType:

Antibody

Datashownislot-specific.Contactusforspecificinformationonotherlots.

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Bpsbioscience/Anti–5–hmC polyclonal antibody (rabbit)/25205/100 µl