Background:

ThisreadytouseNegativeControlreagentcontainsacombinationoffluoresceinandphycoerythrinconjugatedmouseimmunoglobulinmoleculesofIgG1isotype,whichhavebeenselectedonthebasisoftheirbindingcharacteristics:nospecificbindingtohumanintracellularorcellsurfaceantigens,samelowrangeofnonspecificbindingtohumanleukocytesasotherCOMBI-IC-Reagents.LikeallotherCOMBI-IC-reagents,thisreagentshouldbeusedincombinationwithourFIX&PERMCellPermeABIlizationKit(Cat.No.GAS-002)TheseisotypecontrolIgG1aresuitableasnegativecontrolstobeusedincombinationwithCOMBI-ICreagentsforthe:-EnumerationofMyeloidCells-AnalysisofMyeloidDifferentiationStage-EnumerationofB-cellsandPrecursors-EnumerationofT-cellsandPrecursors-AnalysisofLeukemiaCells-AnalysisofImmunodeficiencyStatesResultsmustbeputwithinthecontextofotherdiagnostictestsaswellastheclinicalhistoryofthepatientbyacertifiedprofessionalbeforefinalinterpretation.

Product:

1mlofFITC-andPE-conjugatedVI-APinPBSpH7.2,1%BSA,and0.05%NaN3,approximately50tests.

Applications:

PermeabilizationandStainingProcedure-IncombinationwithourPermeabilizationKitFIX&PERM(Cat.No.GAS-002)intracellularisotypecontrolscanbeeasilystainedincellsUSPensions.-Foreachsampletobeanalyzedadd50µlofwholeblood,bonemarrowormononuclearcellsuspensionina5mltube-Add100µlofReagentA(FixationMedium,storedandusedatroomtemperature)-Incubatefor15minutesatroomtemperature-Add5mlphosphatebufferedsalineandcentrifugecellsfor5minutesat300g-Removesupernatantandaddtocellpellet100µlReagentB(PermeabilizationMedium)and20µlofCOMBI-ICNegativeControlantibodyconjugate.-Vortexatlowspeedfor1-2seconds-Incubatefor15minutesatroomtemperature-Washcellswithphosphatebufferedsalineasdescribedabove-Removesupernatantandresuspendcellsinsheathfluidforimmediateanalysisorresuspendcellsin0.5ml1.0%formaldehydeandstorethemat2-8°Cinthedark.Analyzefixedcellswithin24hours.

Specificity:

VI-APreactswithcalfintestinealkalinephosphataseanddoesnotshowcross-reactivitywithhumanproteins.

Storage:

Nordic-Mubiomonoclonalantibodyreagentscontainoptimalconcentrationsofaffinity-purifiedantibody.Forstabilityreasonsthismonoclonalantibodysolutioncontainssodiumazide.Thesereagentsshouldbestoredat2-8°C(DONOTFREEZE!)andprotectedfromprolongedexposuretolight.Stabilityofthereagent:Pleaserefertotheexpirydateprintedontothevial.Theuseofthereagentaftertheexpirationdateisnotrecommended.

References:

1.J.L.Clarke,W.Watkins,JBiolChem271,10317(1996).
2.R.N.Knibbsetal.,JCellBiol133,911(1996).
3.B.Kniepetal.,JBiochem(Tokyo)119,456(1996).
4.A.J.Wagers,L.M.Stoolman,R.Kannagi,R.Craig,G.S.Kansas,JImmunol159,1917(1997).
5.M.Noguchi,N.Sato,H.Sugimori,K.Mori,K.Oshimi,LeukRes25,847(2001).
6.W.M.Watkins,J.L.Clarke,AdvExpMedBiol491,231(2001).

Caution:

Forprofessionalusersonly.Thisreagentcontainssodiumazide.Toavoidthedevelopmentofhazardousconditions,reagentscontainingazideshouldbedilutedinrunningwaterpriortobediscarded.SimilartotheworkwithotherBIOLOGicalproducts,properhandlingproceduresarerecommended.

Noridic-Mubio是荷兰的一家著名的免疫学试剂生产厂商，因收购An Der Grub Bio Research GmbH(ADG)，增加了自己的产品线：全球最好的固定破膜剂FIX&PERM 。Nordic-Mubio严格遵照ISO9001标准生产高质量的抗体，主要为针对细胞粘附蛋白、核蛋白和骨架蛋白的抗体。同时Nordic-Mubio也是斑马鱼抗体开发的引领者之一，拥有50多种可用于斑马鱼免疫荧光检测的抗体。