Background:
ThisreadytouseNegativeControlreagentcontainspurified,fluoresceinorphycoerythrinconjugatedmouseimmunoglobulinmoleculesofIgG1isotype,whichhavebeenselectedonthebasisoftheirbindingcharacteristics:nospecificbindingtohumancellsurfaceorintracellularantigens,samelowrangeofnonspecificbindingtohumanleukocytesasother
Nordic-MubioReagents.ThisisotypecontrolIgG1issuitableasaNegativeControltobeusedincombinationwithNordic-MUbioreagentsforthe:-EnumerationofMyeloidCells-AnalysisofMyeloidDifferentiationStage-EnumerationofB-cellsandPrecursors-EnumerationofT-cellsandPrecursors-AnalysisofLeukemiaCells-AnalysisofImmunodeficiencyStatesThenegativecontrolreagentpermitstoestimatethedegreeofnon-specificbindingofisotypematchedimmunologbulinstoleukocytesviae.g.Fc-receptors.Itenablestheexperttosetflowcytometricparametersaccordingly.Resultsmustbeputwithinthecontextofotherdiagnostictestsaswellastheclinicalhistoryofthepatientbyacertifiedprofessionalbeforefinalinterpretation.
Product:
2mlofnon-conjugatedVI-APinPBSpH7.2,1%BSA,and0.05%NaN3,approximately100tests.
Applications:
DirectImmunofluorescence(StainingProcedure)Nordic-MUbio
Fluorochromelabeledantibodiesaredesignedforusewitheitherwholebloodorisolatedmononuclearcell(MNC)preparations.Proposedstainingprocedureforwholebloodinshort:-Foreachsampleadd50µlofEDTAanti-coagulatedbloodtoa3-5mltube-Add20µloftheappropriateNordic-MUbiomonoclonalantibodyconjugate-Incubatethetubefor15minutesat4°Coratroomtemperatureinthedark-Add100µlNM-LYSE(Cat.No.GAS-003)toeachtubeandincubatefor10minutesatroomtemperature-Add3-4mlofdestilledwaterandvortex,incubatefor5-10minutesatroomtemperature-Centrifugetubefor5minutesat300g-Aspiratesupernatantandres
USPendpelletin0.3mlofsheathfluid-Analyzeimmediatelyorstoresamplesat2-8°Cinthedarkandanalyzewithin24hoursFor“No-Wash”protocolpleaserefertowww.
Nordicmubio.comProposedstainingprocedureforMNCinshort:-Carefullyadd20µlantibodyconjugateand50-100µlMNCtothebottomofatube-Vortexatlowspeedfor1-2seconds-Incubatefor15-30minutesat2-8°Coratroomtemperature-Centrifugetubesfor5minutesat300g-Removesupernatant,resuspendcellsin2-5mlofphosphatebufferedsaline(PBS)andcentrifugecellsagainfor5minutesat300g-Removesupernatantandresuspendcellsinsheathfluidforimmediateanalysisorresuspendcellsin0.5ml1%formaldehydeandstorethemat2-8°Cinthedark.Analyzefixedcellswithin24hours
Specificity:
ThecloneVI-APreactswithcalfintestinealkalinephosphataseanddoesnotshowcross-reactivitywithhumanproteins.ThesensitivityofVI-APmAbisdeterminedbystainingwell-definedbloodsamplesfromrepresentativedonorswithserial-foldmAbdilutionstoobtainatitrationcurvethatallowsrelatingthemAbconcentrationtothepercentageofstainedcellsandgeometricMFI(meanfluorescenceintensity).Forthispurpose,amAb-concentrationrangeisselectedtoincludeboththesaturationpoint(i.e.themAbdilutionexpectedtobindallepitopesonthetargetcell)andthedetectionthreshold(i.e.themAbdilutionexpectedtorepresenttheleastamountofmAbneededtodetectanidenticalpercentageofcells).Inpractice,50µlofleukocytescontaining10^7cells/mlarestainedwith20µlmAbofvariousdilutionstoobtainatitrationcurveandtoidentifythesaturationpointanddetectionthreshold.Thefinalconcentrationoftheproductisthenadjustedtobeatleast3-foldabovethedetectionthreshold.Inadditionandtocontrollot-to-lotvariation,thegivenlotiscomparedandadjustedtofluorescencestandardswithdefinedintensity.
Storage:
Nordic-MUbiomonoclonalantibodyreagentscontainoptimalconcentrationsofaffinity-purifiedantibody.Forst
ABIlityreasonsthismonoclonalantibodysolutioncontainssodiumazide.Thesereagentsshouldbestoredat2-8°C(DONOTFREEZE!)andprotectedfromprolongedexposuretolight.Stabilityofthereagent:Pleaserefertotheexpirydateprintedontothevial.Theuseofthereagentaftertheexpirationdateisnotrecommended.
Caution:
Forprofessionalusersonly.Thisreagentcontainssodiumazide.Toavoidthedevelopmentofhazardousconditions,reagentscontainingazideshouldbedilutedinrunningwaterpriortobediscarded.Similartotheworkwithother
BIOLOGicalproducts,properhandlingproceduresarerecommended. Nordic的纯化的牛IgG可以在各种免疫测定(包括免疫扩散,免疫电泳,ELISA,Western印迹,斑点免疫结合测定(DIBA),血细胞凝集和细胞结合测定)中用作参考抗原,校准物,包被蛋白和封闭剂。该产品仅用于研究目的,并且用于体外测试,不适用于涉及人或动物的诊断或治疗程序。它可能包含有害成分。请参阅安全数据表(SDS)了解更多信息和正确的处理程序。根据当地法规处理产品残留物。此数据表的准确性尽可能合理,但Nordic-MUbio对本信息中的任何准确性或遗漏概不负责。