Description
CatalogueNumber	17-344
BrandFamily	Upstate
TradeName	Upstate
Description	H2A.XPhosphorylationAssayKit(FlowCytometry)
Overview	PhosphorylationofthehistonevariantH2A.XisarapidandsensitiveresponsetodoublestrandDNAbreaks.TheH2A.XPhosphorylationAssayKit(Flowcytometry)isacellbasedassayformattedforflowcytometricdetectionoflevelsofphosphorylatedhistoneH2A.X. TheH2A.XPhosphorylationAssayKit(Flowcytometry)isacellbasedassayformattedforflowcytometricdetectionoflevelsofphosphorylatedHistoneH2A.X.Cellsareculturedinmicroplates,treatedwithagentsthatinduceDNAdamageorapoptosis,whichstimulatesH2A.Xphosphorylation.CellsarethenfixedandpermeABIlizedinpreparationforstaininganddetection.HistoneH2A.Xphosphorylatedatserine139isdetectedbytheadditionoftheanti-phospho-HistoneH2A.X,FITCconjugate.CellsarethenscannedinaflowcytometertoquantitatethenumberofcellsstainingpositiveforphosphorylatedHistoneH2A.X.

ProductInformation
Components	Anti-phosphoH2A.X,FITCconjugate NormalMouseIgG,FITCconjugate(Cat.#12-487) 16XFixationSolution 10XPermeabilizationSolution 10XWashSolution
Detectionmethod	Fluorescent

StorageandShippingInformation

Applications
Application	TheH2A.XPhosphorylationAssayKit(Flowcytometry)isacellbasedassayformattedforflowcytometricdetectionoflevelsofphosphorylatedHistoneH2A.X.
KeyApplications	FlowCytometry

BIOLOGicalInformation
EntrezGeneNumber	NM_002105.2
EntrezGeneSummary	HistonesarebasicnuclearproteinsthatareresponsIBLeforthenucleosomestructureofthechromosomalfiberineukaryotes.Twomoleculesofeachofthefourcorehistones(H2A,H2B,H3,andH4)formanoctamer,aroundwhichapproximately146bpofDNAiswrappedinrepeatingunits,callednucleosomes.Thelinkerhistone,H1,interactswithlinkerDNAbetweennucleosomesandfunctionsinthecompactionofchromatinintohigherorderstructures.ThisgeneencodesamemberofthehistoneH2Afamily,andgeneratestwotranscriptsthroughtheuseoftheconservedstem-loopterminationmotif,andthepolyAadditionmotif.
GeneSymbol	H2AFX H2A/X H2A.X H2AX H2a/x
Modifications	Phosphorylation
UniProtNumber	P16104
UniProtSummary	FUNCTION:SwissProt:P16104#VarianthistoneH2AwhichreplacesconventionalH2Ainasubsetofnucleosomes.NucleosomeswrapandcompactDNAintochromatin,limitingDNAaccessibilitytothecellularmachinerieswhichrequireDNAasatemplate.Histonestherebyplayacentralroleintranscriptionregulation,DNArepair,DNAreplicationandchromosomalstability.DNAaccessibilityisregulatedviaacomplexsetofpost-translationalmodificationsofhistones,alsocalledhistonecode,andnucleosomeremodeling.Requiredforcheckpoint-mediatedarrestofcellcycleprogressioninresponsetolowdosesofionizingrADIationandforefficientrepairofDNAdoublestrandbreaks(DSBs)specificallywhenmodifiedbyC-terminalphosphorylation. SIZE:143aminoacids;15145Da SUBUNIT:ThenucleosomeisahistoneoctamercontainingtwomoleculeseachofH2A,H2B,H3andH4assembledinoneH3-H4heterotetramerandtwoH2A-H2Bheterodimers.Theoctamerwrapsapproximately147bpofDNA.InteractswithnumerousproteinsrequiredforDNAdamagesignalingandrepairwhenphosphorylatedonSer-140.TheseincludeMDC1,TP53BP1,BRCA1andtheMRNcomplex,composedofMRE11A,RAD50,andNBN.InteractionwiththeMRNcomplexismediatedatleastinpartbyNBN.AlsointeractswithDHX9/NDHIIwhenphosphorylatedonSer-140. SUBCELLULARLOCATION:Nucleus.DEVELOPMENTALSTAGE:SynthesizedinG1aswellasinS-phase. DOMAIN:SwissProt:P16104The[ST]-QmotifconstitutesarecognitionsequenceforkinasesfromthePI3/PI4-kinasefamily. PTM:PhosphorylatedonSer-140(toformgamma-H2AFX)inresponsetoDNAdoublestrandbreaks(DSBs)generatedbyexogenousgenotoxicagentsandbystalledreplicationforks,andmayalsooccurduringmeioticrecombinationeventsandimmunoglobulinclassswitchinginlymphocytes.PhosphorylationcanextenduptoseveralthousandnucleosomesfromtheactualsiteoftheDSBandmaymarkthesurroundingchromatinforrecruitmentofproteinsrequiredforDNAdamagesignalingandrepair.Widespreadphosphorylationmayalsoservetoamplifythedamagesignaloraidrepairofpersistentlesions.PhosphorylationofSer-140inresponsetoionizingradiationismediatedbybothATMandPRKDCwhiledefectsinDNAreplicationinduceSer-140phosphorylationsubsequenttoactivationofATRandPRKDC.DephosphorylationofSer-140byPP2AisrequiredforDNADSBrepair.Inmeiosis,Ser-140phosphorylationmayoccuratsynaptonemalcomplexesduringleptoteneasanATM-dependentresponsetotheformationofprogrammedDSBsbySPO11.Ser-140phosphorylationmaysubsequentlyoccursatunsynapsedregionsofbothautosomesandtheXYbivalentduringzygotene,downstreamofATRandBRCA1activation.Ser-140phosphorylationmayalsoberequiredfortranscriptionalrepressionofunsynapsedchromatinandmeioticsexchromosomeinactivation(MSCI),wherebytheXandYchromosomescondenseinpachytenetoformtheheterochromaticXY-body.Duringimmunoglobulinclassswitchrecombinationinlymphocytes,Ser-140phosphorylationmayoccuratsitesofDNA-recombinationsubsequenttoactivationoftheactivation-inducedcytidinedeaminaseAICDA.&MonoubiquitinationofLys-120byRING1andRNF2/RING2complexgivesaspecifictagforepigenetictranscriptionalrepression(Bysimilarity). SIMILARITY:BelongstothehistoneH2Afamily.

PhysicochemicalInformation

Dimensions

MaterialsInformation

MaterialsInformation

Millipore 提供三种不同的 Immobilon PVDF 膜，是针对不同的蛋白质印迹应用的理想选择。 现在我们还 提供采用按规格裁切的印迹方片膜和两层过滤纸组成的印迹三明治。   与硝酸纤维素膜相比，Immobilon 膜具有多种优点。 它们不易破裂或卷 曲，便于裁剪且在切割过程中不会碎裂。 它们还有低背景、广泛溶剂相容性以及优异的着色能力。 此外，它们可以重复检测。 Immobilon-P 膜（沃比森-供应） ·         0.45μm 孔径 ·         建议用于大多数免疫印迹，特别在蛋 白质大于 20kDa 时 ·         Millipore 的快速免疫检测实验方法不需要膜封闭并缩短洗涤时间，将检测时间至少减少了两个小时，而不致降低灵敏 度或特异性 用于高通量实验室的印迹三明治 ·         当您需要处理多个印迹时，印迹三明治是一种便利省时的选择。 我们的印迹三明 治包含多片 Immobilon-P 膜，其中插入了按规格裁切的多张层析级印迹纸。 转印滤纸 ·         按照大多数免疫印迹尺寸进行裁切的层析级转印滤纸。 Immobilon-PSQ 膜（沃比森-供应） ·         0.2μm 孔径和大的内表面面积 ·         较其它膜具有更高的 蛋白质吸附率和测序得率 ·         建议用于小于 20kDa 的印迹蛋白质 ·         防止小分子量的蛋白质的渗漏 Immobilon-FL 膜（沃比森-供应） ·        针对荧光应用进行优化的转印膜 ·         极低的背景提高了所 有荧光检测实验的灵敏度 ·         与所有常用的荧光探针（所有激发和发射波长）兼容 ·         是多态性检测和化学荧光应用的理想选择