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| Description | |
|---|---|
| CatalogueNumber | HTS004M |
| BrandFamily | Chemicon® |
| TradeName |
|
| 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. |
| BIOLOGicalInformation | |
|---|---|
| Concentration | 2mL,0.5mg/mL |
| EntrezGeneNumber | |
| EntrezGeneSummary | ThisgeneencodesaCXCchemokinereceptorspecificforstromalcell-derivedfactor-1.Theproteinhas7transmembraneregionsandislocatedonthecellsurface.ItactswiththeCD4proteintosupportHIVentryintocellsandisalsohighlyexpressedinbreastcancercells.MutationsinthisgenehavebeenassociatedwithWHIM(warts,hypogammaglobulinemia,infections,andmyelokathexis)syndrome.Alternatetranscriptionalsplicevariants,encodingdifferentisoforms,havebeencharacterized. |
| GeneSymbol |
|
| GPCRClass | A |
| HostCells | Chem-1 |
| ProteinTarget | CXCR4 |
| TargetSub-Family | Chemokine |
| UniProtNumber | |
| 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 |
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