Bpsbioscience/Anti–5–caC polyclonal antibody/25201/20 µg188bio精品生物—专注于实验室精品爆款的电商平台

HostSpecies:Rabbit

Clonality:polyclonal

Background:

Theso-called6thbase,5–hydroxymethylcytosinehasbeendiscoveredin2009.Thefunctionofthisnewmodifiedbaseremainsmostlyunclear,butitiscertainthatthe5–hmCbaseisgeneratedbyenzymaticconversionof5–methylcytosine(5–mC)into5–hydroxymethylcytosinebytheTETfamilyofoxygenases.Earlyreportssuggestedthat5–hmCmayrepresentanintermediateofactivedemethylationinanewpathwaywhichdemethylatesDNA,converting5–mCtocytosine.Recentevidencefuelthehypothesisthat5–hmCrepresentsanintermediate,whichcouldinvolvefurtheroxidationofthehydroxymethylgrouptoaformylorcarboxylgroupfollowedbyeitherdeformylationordecarboxylation.Thecarboxylandformylgroupsof5–formylcytosine(5–fC)and5–carboxylcytosine(5–caC)couldbeenzymaticallyremovedwithoutexcisionofthebase.

Description:

Polyclonalantibodyraisedinrabbitagainst5–Carboxylcytosine(5ca–CMPmonophosphate)conjugatedtoBSA

Synonym(s):

5–Carboxylcytosine

AssayConditions:

Dotblotanalysisusingtheantibodydirectedagainst5-caC
Todemonstratethespecificityoftheantibodyagainst5-caC(Cat.#25201),aDotBlotanalysiswasperformedusingsyntheticoligonucleotidescontainingdifferentmodifiedC-bases(indicatedinred).125and25ngoftherespectiveoligoswereboundtoaStreptavindin-coatedmulti-wellplate.Theantibodywasusedatadilutionof1:1,000.ThebindingofantibodytotheDNAwasmeasuredbyECLchemiluminescence.Figure1showsahighspecificityoftheantibodyforthecarboxylatedcytosine.

Immunofluorescenceassayusingtheantibodydirectedagainst5-caC
293TcellsweretransfectedwitheitherthemouseFLAG-taggedwild-typeTet1(Tet1CD)orthecatalyticallyinactiveFLAG-taggedC-terminaldomainofTet1(Tet1mCD)andstainedwiththeantibodyagainst5-caC(Cat.#25201),diluted1:500,andwithananti-FLAGantibody,followedbyDAPIcounterstaining.

Immunoprecipitationusingtheantibodydirectedagainst5-caC
Immunoprecipitationwasperformedwiththeantibodyagainst5-caC(Cat.#25201)on2µgofJ1ESgenomicDNA,spikedwith1pgofacontrolDNAfragment(approximately700bpfromtheRFP(Ringfingerprotein)gene)containingdifferentcytosinemodifications.ThemCandhmCcontrolDNAwasgeneratedbyPCRwiththecorrespondingnucleotide.ThecaCcontrolfragmentwasobtainedbyinvitromethylationusingM.SssImethyltransferasefollowedbyoxidationwithpurifiedTet2.TheimmunoprecipitatedDNAwassubsequentlyanaysedbyqPCRusingprimersspecificforthecontrolDNAfragmentsandforGAPDH,usedasanegativecontrol.Figure3showstheenrichmentcalculatedastheratiooftherecoveryofthecontrolDNAversustherecoveryoftheGAPDHnegativecontrol.

Concentration:

20µg

Formulation:

PBS(pH7.4),30%glycerol,0.05%sodiumazide

SpeciesReactivity:

CrossReactivity:

Specificfor5–carboxylcytosine(5–caC).Doesnotreactwith5–mc,5–hmc,unmodifiedcytosine,or5–formylcytosine(5–fC)and5–carboxylcytosine(5–caC)

Purification:

ProteinAaffinitypurified

Immunogen:

nucleotide

Format:

Aqueousbuffersolution

Storage/Stability:

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

Application(s):

DB(1:500–1:1000)
IF(1:500)
DNAIP(4µggenomicDNA/IP)

Notes:

Theoptimalantibodyconcentrationshouldbedeterminedbytheend–user.

Warning(s):

Avoidfreeze/thawcycles

ScientificCategory:

Antibodies

ProductType:

Antibody

Datashownislot-specific.Contactusforspecificinformationonotherlots.

对照品标准品，生物试剂盒，gibco胎牛血清，试剂采购网，中国试剂网，细胞因子试剂盒

Bpsbioscience/Anti–5–caC polyclonal antibody/25201/20 µg