Description
CatalogueNumber	HTS055M
BrandFamily	Chemicon®
TradeName	ChemiScreen Chemicon
Description	ChemiSCREEN™MembranePreparationRecombinantHumanCXCR5ChemokineReceptor
Overview	HumanCXCR5
BackgroundInformation	CXCR5(alsoknownasBLR1)isaGPCRthatbindsselectivelytothechemokineCXCL13(alsoknownasBLCandBCA-1)tomediateimmunesystemdevelopmentandfunction(Försteretal.,1996;Gunnetal.,1998).CXCR5cooperateswithCCR7todeterminefunctionalorganizationoflymphoidorganssuchaslymphnodesandPeyer’spatches(Ohletal.,2003).Inaddition,CXCR5mediatesmigrationofBcellsintothefolliclesofthesplenicwhitepulp,thuspermittingantigendepositiononfolliculardendriticcells(Försteretal.,1996;Cinamonetal.,2008).Patientswithrheumatoidarthritis(RA)displayedupregulatedexpressionofCXCR5toagreaterextentthanotherchemokinereceptors,andCXCR5-nullmicesubjectedtoantigen-inducedarthritisdisplayreducedjointdestruction(Schmutzetal,2004;Wengneretal,2007).Therefore,CXCR5representsapotentialtargetfortreatmentofrheumatoidarthritis.Millipore"sCXCR5MembranePreparationforGTPγSAssayisacrudemembranepreparationmadefromtheproprietarystablerecombinantcelllinethatisalsousedforMillipore’sCXCR5CalciumOptimizedCellLine(HTS055C).ThisCXCR5MembranePreparationhasbeenoptimizedforlowbasalactivityinGTPγSbindingassays;thusitisanidealHTStoolforscreeningofcompoundsforagoNISTandantagonistactivities.ThemembranepreparationsexhibitsEC50of1.8nMforCXCL13inaGTPγSbindingassay.

ProductInformation
Format	Membranes
Presentation	Liquidinpackagingbuffer:50mMTrispH7.4,10%glyceroland1%BSAwithnopreservatives. Packagingmethod:Membraneproteinwasadjustedtotheindicatedconcentrationinpackagingbuffer,rapidlyfrozen,andstoredat-80°C.

StorageandShippingInformation
StorageConditions	Storeat–70°C.Productisstableforatleast6monthsfromthedateofreceiptwhenstoredasdirected.Donotfreezeandthaw.

Applications
Application	HumanCXCR5GPCRmembranepreparationforRADIoligandbindingAssays>PγSbinding.
KeyApplications	Radioligandbindingassay GTPγSbinding
ApplicationNotes	RadioligandbindingassayandGTPγSbinding

BIOLOGicalInformation
EntrezGeneNumber	NM_001716
GeneSymbol	CXCR5 BLR1 MDR15
GPCRClass	A
HostCells	Chem-5
ProteinTarget	CXCR5
TargetSub-Family	Chemokine
UniProtNumber	P32302
IncubationConditions	MembranesarepermeABIlizedbyadditionofsaponintoanequalconcentrationbymass,thenmixedwith[35S]-GTPγS(finalconcentrationof0.3nM)in20mMHEPES,pH7.4/100mMNaCl/10mMMgCl2/0.5μMGDPinanonbinding96-wellplate.UnlabeledCXCL13wasaddedtothefinalconcentrationindicatedinFigure1(finalvolume100μL),andincubatedfor30minat30°C.ThebindingreactionistransferredtoanFBfilterplate(MilliporeMAHFB1H)previouslyprewettedwithwater.Theplateiswashed3times(1mLperwellperwash)withcold10mMsodiumphosphate,pH7.4,thendriedandcounted. Onepackagecontainsenoughmembranesforatleast200assays(units),whereoneunitistheamountofmembranethatwillyieldgreaterthan1000cpmspecificCXCL13-stimulated[35S]-GTPγSbinding.

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