Bpsbioscience/Anti–AML1–ETO polyclonal antibody/25208/100 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:

TheAML1(RUNX1)(UniProtKB/Swiss–ProtentryQ01196)–ETO(RUNX1T1)(UniProtKB/Swiss–ProtentryQ06455)fusionproteinarisesduetoatranslocationbetweenchromosome8and22(t(8;21)(q22;q22)).Thistranslocationisoneofthemostfrequentkaryotypicabnormalitiesobservedinacutemyeloidleukaemia.Itproducesachimericalgenemadeupofthe5’–regionofAML1andthe3’–regionofETO.Thechimericalproteinisthoughttoassociatewiththenuclearcorepressor/histonedeacetylasecomplextoblockhematopoieticdifferentiation.

Description:

PolyclonalantibodyraisedinrabbitagainsttheAML1–ETOfusionproteinusingaKLH–conjugatedsyntheticpeptide.

Synonym(s):

RUNX1–RUNX1T1fusionprotein

AssayConditions:

ChIPresultsobtainedwiththeantibodydirectedagainstAML1-ETO
ChIPassayswereperformedusingKasumi-1cells,theantibodyagainstAML1-ETO(Cat.#25208)andoptimizedprimerpairsforqPCR.Shearedchromatinfrom1.25millioncellsand4µlofantibodywereusedperChIPexperiment.QPCRwasperformedusingprimersspecificfortheFUT7,NFE2andOGG1genes.Figure1showstheoccupancy,calculatedastheratio+control/backgroundforwhichthepromoteroftheH2Bgenewasused.

ChIP-seqresultsobtainedwiththeantibodydirectedagainstAML1-ETO
ChIPwasperformedasdescribedabove.TheimmunoprecipitatedDNAof6ChIPassayswaspooledandanalyzedwithanIlluminaGenomeAnalyzer.Librarypreparation,clustergenerationandsequencingwereperformedaccordingtothemanufacturersinstructions.The32bptagswerealignedtothehumanreferencegenome(hg18)usingtheELANDalgorithm.Figure2showstheresultsofthecompletechromosome3andthreegenomicregionssurroundingtheOGG1,FUT7andNFE2genes,respectively.ThepositionofthePCRampliconisindicatedwithanarrow.

Determinationoftheantibodytiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstAML1-ETO(Cat.#25208).Theplateswerecoatedwiththepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure3),thetiteroftheantibodywasestimatedtobe1:32,750.

WesternblotanalysisusingtheantibodydirectedagainstAML1-ETO
NuclearextractsofSKNO-1cells(15µg)wereanalyzedbyWesternblotusingtheantibodyagainstAML1-ETO(Cat.#25208)diluted1:1,000inTBS-Tweencontaining5%skimmedmilk.TheMarker(inkDa)isshownontheleft,thepositionoftheproteinofinterestisindicatedontheright.

Concentration:

100µl

Formulation:

Wholeantiserumfromrabbitcontaining0.05%azide

SpeciesReactivity:

Human

Purification:

Wholeantiserum

Immunogen:

syntheticpeptide

Format:

aqueoussolution

Storage/Stability:

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

Application(s):

ChIP/ChIP–seq(4µl/ChIP)
ELISA(1:500)
WB(1:1000)

Notes:

TheoptimalantibodyamountperIPshouldbedeterminedbytheend–user.Werecommendtesting1–5µgperIP

Warning(s):

Avoidfreeze/thawcycles

ScientificCategory:

Antibodies

ProductType:

Antibody

Datashownislot-specific.Contactusforspecificinformationonotherlots.

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Bpsbioscience/Anti–AML1–ETO polyclonal antibody/25208/100 µl