FIX&PERM® Solution A (Fix) (CE)

货号：	GAS-002A-1
供应商：	广州柏赛柯生物
数量：	请咨询
规格：	1x100ml

FIX&PERM®SolutionA(Fix)(CE)

具体报价请参考官网：http://www.nordicmubio.com
或咨询以下联系方式：

客服热线：020-39341005

联系客服QQ:1195082744

Cataloguenumber: GAS-002A-1

ProductType	OtherReagents
Units	1x100ml
Application	Flowcytometry

Background
Flowcytometricanalyseswithmonoclonalantibodiesweresofarmainlyrestrictedtocellsurfacemolecules.Intracellularstructuressuchascytoplasmicornuclearenzymes,oncoproteins,cytokines,immunoglobulinsetc.werelargelyexcludedfromsuchstudies.Alsoexcludedfromflowcytometricstudieswerecytoplasmiclocalizationsofwell-establishedmembranemoleculeslikeCD3andCD22,which,intheircytoplasmicform,arethemostreliablelineagemarkersinundifferentiatedleukemia.WiththeFIX&PERM®Kitflowcytometricanalysisofintracellularantigenshasbecomeaseasyassurfaceantigenstudies.Theonlyprerequisiteistheavailabilityofsuitableantibodyconjugates.MostoftheavailablemonoclonalantibodyconjugatescanbeusedwiththeFIX&PERM®Kit,somedeterminantsaresensitive,however,tothefixationstepinvolved.Thisandtheoptimalfixationtimehavetobetestedforeachreagent.

Product
FIX&PERM®SolutionAconstitutesthefixationbufferintheFIX&PERM®Kit.ThisFIX&PERM®CellFixationandPermeabilizationKitcontains2reagents:FixationMedium(ReagentA)andPermeabilizationMedium(ReagentB).ItisintendedforfirstfixingcellsinsuspensionwithReagentAandthenpermeabilizingthecellmembraneswithReagentB.Thisproceduregivesantibodiesaccesstointracellularstructuresandleavesthemorphologicalscattercharacteristicsofcellsintact.Specificformulationsreducebackgroundstainingandallowsimultaneousadditionofpermeabilizationmediumandfluorochromelabeledantibodies.FIX&PERM®issuitablefortheanalysisofnormalandmalignantleukocytepopulationsderivedfromvarioushumanbiologicalsamples(blood,bonemarrowandothers)usingflowcytometry.Resultsmustbeputwithinthecontextofotherdiagnostictestsaswellastheclinicalhistoryofthepatientbyacertifiedprofessionalbeforefinalinterpretation.

Format:1x100mlvialofFixationMedium(ReagentA).FIX&PERM®Reagentsaredesignedforusewithallcommerciallyavailableflowcytometers.Alignmentandcompensationshouldbeperformedaccordingtomanufacturer´sinstructions.

ThequalityofeachFIX&PERM®Lotisdeterminedbyfixationandpermeabilizationofwelldefinedbloodsamplesfromrepresentativedonorsandsubsequentcomparisonofforwardandsidescattercharacteristicsofobtainedleukocytes,aswellasimmunostainingefficiencyforseveralmembranousandcytoplasmicantigens

Applications
Biologicalfluids(blood,bonemarrow,andothers)mustbecollectedundersterileconditions.AnticoagulationwithEDTAorheparinisrecommended.Thesamplesshouldbestoredatroomtemperatureuntilused.Foroptimalresults,samplesshouldbeprocessedandanalyzedwithin24hours.Sampleswithhighnumbersofnon-viablecellsmightcausefalseresults,suchcasesrequiredeterminationofcellviabilitywithe.g.propidiumiodide.Allbiologicalsampleshavetobehandledwithcaution.Alwaysconsiderthemaspotentiallyinfective.Useappropriateprecautionssuchasgloves,lab-coat,etc.

Fixation,permeabilizationandstainingprocedure(whencombinedwithReagentB/GAS-002B):

•Foreachsampletobeanalyzedadd50µlofwholeblood,bonemarrowormononuclearcellsuspensionina5mltube
•Add100µlofReagentA(FixationMedium,storedandusedatroomtemperature)
•Incubatefor15minutesatroomtemperature
•Add5mlphosphatebufferedsalineandcentrifugecellsfor5minutesat300g
•Removesupernatantandaddtocellpellet100µlReagentB(PermeabilizationMedium)and20µloftheappropriateNordic-MUbiomonoclonalantibodyconjugate
•Vortexatlowspeedfor1-2seconds
•Incubatefor15minutesatroomtemperature
•Washcellswithphosphatebufferedsalineasdescribedabove
•Removesupernatantandresuspendcellsinsheathfluidforimmediateanalysisorresuspendcellsin0.5ml1.0%formaldehydeandstorethemat2-8°Cinthedark.Analyzefixedcellswithin24hours.

Comments:Specialcases(dilutedbonemarrowsamples,othersamplescontaininglowsolubleprotein)mightbenefitfromreplenishmentwithplasmacomponentsbeforetheFIX&PERM®treatmentinordertocreateamilieu,whichmorecloselyresemblesthestuationinanti-coagulatedblood.ForthatpuposeadditionofIgGpreparations(e.g.BeriglobulinP,ZLBBehring,finalconcentration10mg/ml)andhumanserumalbumin(e.g.humanalbumin“Behring”20%-infusionsolution,finalconcentration40mg/ml)isrecommended.

Limitationsofthetechnique
Flowcytometryshouldbeperformedbyprofessionalusersonly.Improperalignmentoftheflowcytometer,inaccuratecompensationoffluorescenceleakingintootherchannelsaswellasincorrectpositioningofregionsmayleadtofalseresults.
Lysisofredcellsmightbeimpossibleforvariousreasons.Insuchinstancesitisrecommendedtoisolatemononuclearcells(MNC)viadensitygradientcentrifugationpriortostaining.

Resultswillbecorrectandreproducibleaslongastheproceduresusedrespectthetechnicalrecommendationsandobeygoodlaboratorypractice.

TheFIX&PERM®solutionsareprovidedinaconcentrationthatwillallowtofixandpermeabilizehumanhematopoieticcells.Itisthereforestronglyrecommendedtosticktotheworkingprotocolintermsofconcentrationandvolumeregardingcellsandantibody.ThepropertiesofFIX&PERM®havebeendeterminedusingEDTAanti-coagulatedperipheralblood.

Warranty
Theproductssoldhereunderarewarrantedonlytoconformtothequantityandcontentsstatedonthelabelatthetimeofdeliverytothecustomer.Therearenowarranties,expressedorimplied,thatextendbeyondthedescriptiononthelabeloftheproduct.Nordic-MUbio`ssoleliabilityislimitedtoeitherreplacementoftheproductsorrefundofthepurchaseprice.Nordic-MUbioisnotliableforpropertydamage,personalinjury,oreconomiclosscausedbytheproduct.

Storage
FIX&PERM®CellFixationandPermeabilizationKitreagentsshouldbestoredandusedatroomtemperature.Donotfreeze.Stabilityofthereagent:Pleaserefertotheexpirydateprintedontothevial.Theuseofthereagentaftertheexpirationdateisnotrecommended.Ifreagentsarestoredunderanyconditionsotherthanthosespecified,theconditionsmustbeverifiedbytheuser.Donotusereagentsifaprecipitateshouldformordiscolorationoccurs.Ifunexpectedresultsareobtainedwhichcannotbeattributedtodifferencesinlaboratoryprocedures,pleasecontactus.

References
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-Groeneveld,K.,teMarvelde,J.G.,vandenBeemd,M.W.,Hooijkaas,H.&vanDongen,J.J.(1996)Leukemia10,1383-9.
-Haranaga,S.,Yamaguchi,H.,Friedman,H.,Izumi,S.,&Yamamoto,Y.(2001)InfectImmun69,7753-9.
-Hegazy,A.N.&Klein,C.(2008)Leukemia22,2070-9.
-Kappelmayer,J.,Gratama,J.W.,Karaszi,E.,Menendez,P.,Ciudad,J.,Rivas,R.&Orfao,A.(2000)JImmunolMethods242,53-65.
-Kline,M.P.,Rajkumar,S.V.,Timm,M.M.,Kimlinger,T.K.,Haug,J.L.,Lust,J.A.,Greipp,P.R.&Kumar,S.(2007)Leukemia21,1549-60
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Caution
Forprofessionalusersonly.ReagentAofFIX&PERM®CellFixationandPermeabilizationKitcontainsfomaldehydeandislabelled:Harmful.Formaldehydeistoxic,allergenicandasuspectedcarcinogen.Neverpipettebymouthandavoidcontactwitheyes,skinandclothing.Properhandlingproceduresarerecommended.Asamainrule,personsunder18yearsofagearenotallowedtoworkwiththisproduct.Usersmustbecarefullyinstructedintheproperworkingprocedure,thedangerouspropertiesoftheproductandthenecessarysafetyinstructions.PleaserefertotheMaterialSafetyDataSheet(MSDS)foradditionalinformation.Disposeproductremaindersaccordingtolocalregulations.

Figure1:Flowcytrometricanalysisofnormalbonemarrow(BM)afterfixationandpemeabilizationusingGAS-002,followedbyimmunostainingforlactoferrinandMPO-C2.

Figure2:Flowcytrometricanalysisofbonemarrow(BM)fromaleukemiapatientafterfixationandpemeabilizationusingGAS-002,followedbyimmunostainingforlactoferrinandMPO-C2.

Figure3:ImmunostainingwithNordic-MUbioCD22-PEconjugateofundifferentiatedleukemiacellsofB-ALLtype.

Figure4:ImmunostainingwithNordic-MUbioCD3-PEconjugateofsurfaceCD3-negativeundifferentiatedleukemiacellsofT-ALLtype.

Figure5:ImmunostainingwithNordic-MUbioantiMPO-FITCconjugateofundifferentiatedleukemiacellsofAMLtype.

Figure6:ForwardandsidescatterpatternofanormalbloodsampletreatedwithGAS-002.

Figure7:DoublelabelingofanormalbloodsampletreatedwithGAS-002,andimmunostainedforLactoferrin(PE)andMPO-C2(FITC).

Figure8;DoublelabelingofanormalbloodsampletreatedwithGAS-002,andimmunostainedforCD3(PE)andMPO-C2(FITC).

Figure9:DoublelabelingofanormalbloodsampletreatedwithGAS-002,andimmunostainedforLactoferrin(PE)andLysozyme(FITC).

温馨提示：不可用于临床治疗。

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FIX&PERM® Solution A (Fix) (CE)

FIX&PERM®SolutionA(Fix)(CE)

具体报价请参考官网：http://www.nordicmubio.com或咨询以下联系方式：客服热线：020-39341005 联系客服QQ:1195082744

具体报价请参考官网：http://www.nordicmubio.com
或咨询以下联系方式：

客服热线：020-39341005

联系客服QQ:1195082744