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 Immobilon -P转印膜   PVDF膜 ·适用于免疫印迹、斑点印迹和其他蛋白质印迹的最佳PVDF膜；·Western Blotting、吸附分析、氨基酸分析、N端蛋白质测序、Dot/Slot blotting、糖蛋白显色和脂多糖分析；·与他相容的染液/染色剂：考马斯亮蓝染色剂、酰胺黑、印度墨水、丽春红、胶体金、CPTS、甲苯胺蓝、透射显影和Sypro Ruby染料. 产品名称Immobilon-P说    明应用于>20KD蛋白质的Western, dot/slot blotting和其他印迹孔    径0.45um尺寸cm×cm26.5×375数    量1卷目录编号IPVH00010 ――――――――――――――――――――――――――――――――――――――― Immobilon - NC膜，混合纤维素酯（Millipore） Immobilon-NC HATF由混合纤维素酯构成，不含细胞生长期间影响细胞完整的表面活性剂。使用这些膜可从事菌落转移实验。Immobilon-NC HAHY由混合纤维素酯构成，但含有转印过程期间能改善可湿性便于操作的表面活性剂。膜的孔径是0.45μm。用于Southern blotting，Northern Blotting，Western blotting，Colony/plaque Lifts，Dot/slot blots，吸附分析，细胞培养.。 产品名称Immobilon - NC说    明应用于Southern杂交， Northern杂交，免疫印迹，结合实验孔    径0.45um尺寸cm×cm33×300数    量1卷目录编号HATF00010