Description
CatalogueNumber	HTS004M
BrandFamily	Chemicon®
TradeName	ChemiScreen Chemicon
Description	ChemiSCREEN™CXCR4MembranePreparation
Overview	Full-lengthhumanCXCR4CDNA
BackgroundInformation	ThechemokineSDF-1aanditsGPCRreceptorCXCR4haveaone-to-onespecificitythatisuniqueamongchemokinesandtheirreceptors.SDF-1abindstoCXCR4expressedonhematopoieticandlymphopoieticcells,anddirectstheirtraffickingtoandretentioninhemato-andlymphatopoieticorgansandsitesofinflammation(Kuciaetal.,2004).CXCR4isexpressedonseveraltumorcelllineages,andmightberesponsIBLeformetastasistositesofSDF-1aexpression,suchasboneandlymphnodes(Mulleretal.,2001).Inaddition,CXCR4isacoreceptorfortheHIVenvelopeglycoproteingp120(Fengetal.,1996).SmallmoleculeantagoNISTsofCXCR4havebeendevelopedandshowntoinhibitinfectivityofT-tropicHIVstrainsandtoimpairgrowthofbraintumors(Arakakietal.,1999;Rubinetal.,2003).Chemicon"sCXCR4membranepreparationsarecrudemembranepreparationsmadefromourproprietarystablerecombinantcelllinestoensurehigh-levelofGPCRsurfaceexpression;thus,theyareidealtoolsforscreeningforantagonistsofinteractionsbetweenCXCR4andSDF-1a.RADIolabeledSDF-1abindstotheCXCR4membraneswithaKdof0.9nM.With0.04-0.13nM125I-labeledSDF-1a,anapproximately3-foldwindowofspecificbindingisobtained.

ProductInformation
Format	Membranes
Presentation	Packagingbuffer:50mMTrispH7.4,10%glyceroland1%BSA Packagingmethod:Membranesproteinwereadjustedto0.5mg/mlin1mlpackagingbuffer,rapidlyfrozen,andstoredat-80°C.

StorageandShippingInformation
StorageConditions	Maintainfrozenat-70°Cforupto2years.Donotfreezeandthaw.

Applications
Application	HumanCXCR4GPCRmembranepreparationforRadioligandbindingAssays>PgammaSbinding.
KeyApplications	Radioligandbindingassay GTPγSbinding
ApplicationNotes	RadioligandbindingassayandGTPgammaSbinding.

BIOLOGicalInformation
Concentration	2mL,0.5mg/mL
EntrezGeneNumber	NM_001008540.1 NM_003467.2
EntrezGeneSummary	ThisgeneencodesaCXCchemokinereceptorspecificforstromalcell-derivedfactor-1.Theproteinhas7transmembraneregionsandislocatedonthecellsurface.ItactswiththeCD4proteintosupportHIVentryintocellsandisalsohighlyexpressedinbreastcancercells.MutationsinthisgenehavebeenassociatedwithWHIM(warts,hypogammaglobulinemia,infections,andmyelokathexis)syndrome.Alternatetranscriptionalsplicevariants,encodingdifferentisoforms,havebeencharacterized.
GeneSymbol	CXCR4 WHIM NPY3R NPYY3R FB22 LAP3 Fusin CXCR-4 LCR1 NPYR HM89 fusin HSY3RR CXC-R4 NPYY3 NPYRL D2S201E CD184 LESTR
GPCRClass	A
HostCells	Chem-1
ProteinTarget	CXCR4
TargetSub-Family	Chemokine
UniProtNumber	P61073
UniProtSummary	FUNCTION:SwissProt:P61073#ReceptorfortheC-X-CchemokineCXCL12/SDF-1.Transducesasignalbyincreasingtheintracellularcalciumionslevel.Involvedinhaematopoiesisandincardiacventricularseptumformation.Playsalsoanessentialroleinvascularizationofthegastrointestinaltract,probablybyregulatingvascularbranchingand/orremodelingprocessesinendothelialcells.Couldbeinvolvedincerebellardevelopment.IntheCNS,couldmediatehippocampal-neuronsurvival.Actsasacoreceptor(CD4beingtheprimaryreceptor)forHIV-1X4isolatesandasaprimaryreceptorforsomeHIV-2isolates.PromotesEnv-mediatedfusionofthevirus. SIZE:352aminoacids;39746Da SUBUNIT:InteractswithHIV-1surfaceproteingp120andTat. SUBCELLULARLOCATION:Cellmembrane;Multi-passmembraneprotein. TISSUESPECIFICITY:Expressedinnumeroustissues,suchasperipheralbloodleukocytes,spleen,thymus,spinalcord,heart,placenta,lung,liver,skeletalmuscle,kidney,pancreas,cerebellum,cerebralcortexandmedulla(inmicrogliaaswellasinastrocytes),brainmicrovascular,coronaryarteryandumbilicalcordendothelialcells.Isoform1ispredominantinalltissuestested. DOMAIN:SwissProt:P61073Theamino-terminusiscriticalforligandbinding.ResiduesinallfourextracellularregionscontributetoHIV-1coreceptoractivity. PTM:Sulfated. DISEASE:SwissProt:P61073#DefectsinCXCR4areacauseofWHIMsyndrome[MIM:193670];alsocalledwarts,hypogammaglobulinemia,infections,andmyelokathexis.WHIMsyndromeisanimmunodeficiencydiseasecharacterizedbyneutropenia,hypogammaglobulinemiaandextensivehumanpapillomavirus(HPV)infection.Despitetheperipheralneutropenia,bonemarrowaspiratesfromaffectedindividualscontainabundantmaturemyeloidcells,aconditiontermedmyelokathexis. SIMILARITY:BelongstotheG-proteincoupledreceptor1family.
IncubationConditions	RECOMMENDEDASSAYCONDITIONS:Membranesaremixedwithradioactiveligandandunlabeledcompetitor(seeFigures1and2forconcentrationstested)inbindingbufferinanonbinding96-wellplate,andincubatedfor1-2h.Priortofiltration,aGF/C96-wellfilterplateiscoatedwith0.33%polyethyleneiminefor30min,thenwashedwith50mMHEPES,pH7.4,0.5%BSA.Bindingreactionistransferredtothefilterplate,andwashed3times(1mLperwellperwash)withWashBuffer.Theplateisdriedandcounted. Bindingbuffer:50mMHepes,pH7.4,5mMMgCl2,1mMCaCl2,0.2%BSA,filteredandstoredat4°C Radioligand:[125I]SDF-1alpha(Perkinelmer#:NEX346) WashBuffer:50mMHepes,pH7.4,500mMNaCl,0.1%BSA,filteredandstoredat4°C. Onevialcontainsenoughmembranesforatleast200assays(units),whereanunitistheamountofmembranethatwillyieldgreaterthan4-foldsignal:backgroundwith125I-labeledSDF-1alphaat0.13nM.

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