Bpsbioscience/Anti–H3 pan polyclonal antibody/25235/100 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:

Histonesarethemainconstituentsoftheproteinpartofchromosomesofeukaryoticcells.Theyarerichintheaminoacidsarginineandlysineandhavebeengreatlyconservedduringevolution.HistonespacktheDNAintotightmassesofchromatin.TwocorehistonesofeachclassH2A,H2B,H3andH4assembleandarewrappedby146basepairsofDNAtoformoneoctamericnucleosome.Histonesplayacentralroleintheregulationoftranscription,DNArepair,DNAreplicationandchromosomalstability.Thesedifferentfunctionsareestablishedviaacomplexsetofpost–translationalmodificationswhicheitherdirectlyorindirectlyalterchromatinstructureandDNAaccessibilitytofacilitatetranscriptionalactivationorrepressionorothernuclearprocesses.

Description:

PolyclonalantibodyraisedinrabbitagainsttwoKLH–conjugatedsyntheticpeptidescontaininganunmodifiedsequencefromthecentralpartandfromtheC–terminusofhistoneH3,respectively.

AssayConditions:

Determinationofthetiter
Todeterminethetiteroftheantibody,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstH3pan(Cat.#25235).Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:900,000.

CrossreactivitytestusingtheantibodydirectedagainstH3pan
ADotBlotanalysiswasperformedtotestthecrossreactivityoftheantibodyagainstH3pan(Cat.#25235)withthepeptidesusedforimmunizationoftherabbitandotherpeptidescontainingdifferentmodificationsorunmodifiedsequencesofhistoneH3.Onehundredto0.2pmoloftherespectivepeptideswerespottedonamembrane.Theantibodywasusedatadilutionof1:1,000.Figure2showsahighspecificityoftheantibodyforoneoftheunmodifiedpeptides.

WesternblotanalysisusingtheantibodydirectedagainstH3pan
HistoneextractsofHeLacells(15µg)wereanalyzedbyWesternblotusingtheantibodyagainstH3pan(Cat.#25235)diluted1:250inTBS-Tweencontaining5%skimmedmilk(lane1).Lane2showsacoomassiebluestainingofthegel.Thepositionoftheproteinofinterestisindicatedontheleft;theMarker(inkDa)isshownontheright.

ChIPresultsobtainedwiththeantibodydirectedagainstH3pan
ChIPassayswereperformedusinghumanHeLacells,theantibodyagainstH3pan(Cat.#25235)andoptimizedPCRprimersetsforqPCR.ChIPwasperformedonshearedchromatinfrom10,000cellsperIP.Atitrationoftheantibodyconsistingof1,5,and10µlperChIPexperimentwasanalyzed.IgG(5µg/IP)wasusedasnegativeIPcontrol.QPCRwasperformedwithprimersfortheGAPDHpromoter,MYOD,andtheheterochromatinmarkerSat2.Figure4showstherecovery,expressedasa%ofinput(therelativeamountofimmunoprecipitatedDNAcomparedtoinputDNAafterqPCRanalysis).

Concentration:

100µl

Formulation:

Wholeantiserumfromrabbitcontaining0.05%azide

SpeciesReactivity:

Human

Purification:

Wholeserum

Immunogen:

syntheticpeptide

Format:

aqueoussolution

Storage/Stability:

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

Application(s):

ChIP(1–5µl/ChIP)
ELISA(1:100–1:1000)
DB(1:1000)
WB(1:250)

Notes:

TheoptimalantibodyamountperIPshouldbedeterminedbytheend–user.Werecommendtesting1–5µgperIP

Warning(s):

Avoidfreeze/thawcycles

ScientificCategory:

Antibodies

ProductType:

Antibody

Datashownislot-specific.Contactusforspecificinformationonotherlots.

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Bpsbioscience/Anti–H3 pan polyclonal antibody/25235/100 µl