ApplicationNotes | ChromatinImmunoprecipitation(ChIP)Analysis:Arepresentativelot(3μLperChIP)detectedhistoneH4Arg3citrullination(H4Cit3)enrichmentatthepS2genepromoterinMCF-7cellsthathadbeentreatedwithβ-estradiolforonehouraftera3-dayserum-starvationperiod.
ImmunocytochemistryAnalysis:A1:500dilutiondetectedfromarepresentativelotdetectedenhancedhistoneH4Arg3citrullinationinionomycin-stimulatedprimaryhumanneutrophils(CourtesyofKerriMowen,Ph.D.,PadlockTherapeutics).
ImmunohistochemistryAnalysis:A1:250dilutionfromarepresentativelotdetectedenhancedhistoneH4Arg3citrullination(H4Cit3)in4%paraformaldehyde-fixed,decalcified,paraffin-embeddedhindpawsectionsfromwild-type,butnotPad4-knockoutmice5dayspostimmunizationwitharthritogenicserumfromK/BxNmice(CourtesyofKerriMowen,Ph.D.,PadlockTherapeutics).
ChromatinImmunoprecipitation(ChIP)Analysis:Arepresentativelotdetectedatime-dependenthistoneH4Arg3citrullination(H4Cit3)enrichmentatthepS2genepromoterinMCF-7cellsfollowingestradioltreatment(Wang,Y.,etal.(2004).Science.306(5694):279-283).
ImmunocytochemistryAnalysis:ArepresentativelotdetectedenhancednuclearH4/H2AAr3citrullinationinHEK293TandU2OScellsuponexogenousPAD4expressionorDNAdamageinduction(Tanikawa,C.,etal.(2012).Nat.Commun.3:676).
ImmunocytochemistryAnalysis:ArepresentativelotdetectedArg3-citrullinatedhistoneH4(H4Cit3)in3.8%paraformaldehyde-fixed,2%NP-40/1%TritonX-100-permeABIlizedcasein-elicitedperitonealneutrophilscellsfromwild-type,butnotPad4-knockoutmice(Hemmers,S.,etal.(2011).PLoSOne.6(7):e22043).
ImmunocytochemistryAnalysis:ArepresentativelotdetectedArg3-citrullinatedhistoneH4(H4Cit3)in3.8%paraformaldehyde-fixed,2%NP-40/1%TritonX-100-permeabilizedinfiltratinglungleukocytesisolatedfromwild-type,butnotPad4-knockoutmicefollowinginfluenzainfection(Hemmers,S.,etal.(2011).PLoSOne.6(7):e22043).
ImmunocytochemistryAnalysis:ArepresentativelotdetectedArg3-citrullinatedhistoneH4(H4Cit3)in4%paraformaldehyde-fixed,1%NP-40/0.5%TritonX-100-permeabilizedperipheralbloodcellsfromwild-type,butnotPad4-knockoutmice5dayspostimmunizationwitharthritogenicserumfromK/BxNmice(Rohrbach,A.S.,etal.(2012).ArthritisRes.Ther.14(3):R104).
ImmunocytochemistryAnalysis:ArepresentativelotdetectedArg3-citrullinatedhistoneH4(H4Cit3)insynovialcellsandextracellulartraps(NETs)flushedoutoftheanklejointsofwild-type,butnotPad4-knockoutmice5dayspostimmunizationwitharthritogenicserumfromK/BxNmice(Rohrbach,A.S.,etal.(2012).ArthritisRes.Ther.14(3):R104).
ImmunocytochemistryAnalysis:Arepresentativelotdetectedcalciumionophore-inducedhistoneH4(H4Cit3)Arg3citrullinationin3.7%paraformaldehyde-fixed,0.2%NP-40/0.1%TritonX-100-permeabilizedHL-60cells(Wang,Y.,etal.(2004).Science.306(5694):279-283). ImmunohistochemistryAnalysis:Arepresentativelotdetectedgammairradiation-inducednuclearH4/H2AAr3citrullinationinthymustissuesectionsfromwild-type,butnotPad4-knockoutmice(Tanikawa,C.,etal.(2012).Nat.Commun.3:676).
ImmunohistochemistryAnalysis:ArepresentativelotdetectedenhancedhistoneH4Arg3citrullination(H4Cit3)in4%paraformaldehyde-fixed,decalcified,paraffin-embeddedhindpawsectionsfromwild-type,butnotPad4-knockoutmice10dayspostimmunizationwitharthritogenicserumfromK/BxNmice(Rohrbach,A.S.,etal.(2012).ArthritisRes.Ther.14(3):R104).
WesternBlottingAnalysis:ArepresentativelotdetectedPAD4-catalyzedArg3citrullinationofrecombinanthistoneH4andH2A,aswellasDNAdamage-inducedcellularH4andH2AAr3citrullination(Tanikawa,C.,etal.(2012).Nat.Commun.3:676).
WesternBlottingAnalysis:Arepresentativelotdetectedcalciumionophore-inducedhistoneH4Arg3citrullination(H4Cit3)inHL-60cells.CellularPAD4knockdownbyshRNAtreatmentgreatlydiminishedionophor-inducedH4Cit3,whileantibodyblockingwiththeimmunogenpeptidepreventedtargetbanddetection(Wang,Y.,etal.(2004).Science.306(5694):279-283). |