Goat Anti-Mouse α-glucosidase

Goat Anti-Mouse α-glucosidase

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2018-09-27 10:00 873阅读次数

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GoatAnti-Mouseα-glucosidaseStorage:2-8°CPackagesize:96determinationsPRINCIPLEOFTHEMETHODTheα-glucosidasekitisasolidphasephasesandwichenzymelinkedimmunosorbentassay(ELISA).Samples,includingstandardsofknownα-glucosidaseconcentrationsandunknownsarepipettedintothesewells.Duringthefirstincubation,theα-glucosidaseantigenandabiotinylatedmonoclonalantibodyspecificforα-glucosidasearesimultaneouslyincubated.Afterwashing,theenzyme(streptavidin-peroxydase)isadded.Afterincubationandwashingtoremovealltheunboundenzyme,asubstratesolutionwhichisactingontheboundenzymeisaddedtoinduceacolouredreactionproduct.Theintensityofthiscolouredproductisdirectlyproportionaltotheconcentrationofα-glucosidasepresentinthesamples.REAGENTSPROVIDEDANDRECONSTTTUTIONREAGENTS（Storeat2-8℃）1×96WELLS0.5×96WELLSRECONSTTTUTION96/48-wellsmicrotiterplates10.5Ready-to-usePlastivcover21Ready-to-useStandard:400nmol/L1Vials(0.6ml)0.5Vials(0.3ml)Seereagentspreparationonpage3Blankcontrol1Vials(1.0ml)1Vials(0.5ml)Ready-to-useStandardDiluent1Vials(4.0ml)1Vials(2.0ml)Ready-to-useBiotinylatedanti-α-glucosidase1Vials(6.0ml)1Vials(3.0ml)Ready-to-useStreptavidin-HRP1Vials(8.0ml)1Vials(4.0ml)Ready-to-useWashingBuffer1Vials(20ml)1Vials(10ml)50×concentrateSubstrateA1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSubstrateB1Vials（6.0ml)1Vials(3.0ml)Ready-to-useStoppingSolution1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSampleDiluent1Vials(12ml)1Vials(6.0ml)Ready-to-useGoatAnti-Mouseα-glucosidaseMATERIALREQUIREDBUTNOTPROVIDED?Distilledwater?Pipettes:10ul、50ul、100ul、200ul、1000ul。?Vortexmixerandmagneticstirrer.SAFETY?Forresearchuseonly?AvoidanyskincontactwithH2SO4andTMB.Incaseofcontact,washthoroughlywater.?Donoteat,drink,smokeorapplycosmeticswherekitreagentsareused.?Donotpipettebymouth.PROCEDURALNOTES/LAB.QUALITYCONTROL?Whennotinuse,kitcomponents首ldbestoredrefrigeratedorfrozenasindicatedonvialsorbottles.Allreagents首ldbewarmedtoroomtemperaturebeforeuse.Lyophilizedstandards首ldbediscardedafteruse.?Oncethedesirednumberofstripshasbeenremoved,immediatelyresealthebagtoprotecttheremainingstripsfromedterioration.?Coverorcapallreagentswhennotinuse.?Donotmisorinterchangereagentsbetweendifferentlots.?Donotusereagentsbeyondtheexpirationdateofthekit.?Useacleandisposableplasticpipettetipforeachreagent,standard,orspecimenadditioninordertoavoidcross-contamination,forthedispensingofH2SO4andsubstratesolution,avoidpipetteswithmetalparts.?Useacleanplasticcontainertopreparethewashingsolution.?Thoroughlymixthereagentsandsamplesbeforeusebyagitationorswir领.?Allresidualwashingliquidmustbedrainedfromthewellsbyefficientaspirationorbydecantationfollowedbytappingtheplateforcefullyonabsorbentpaper.Neverinsertabsorbentpaperdirectlyintothewells.?TheTMBsolutionislightsensitive.Avoidprolongedexposuretolight,also,avoidcontactoftheTMBsolutionwithmetaltopreventcolourdevelopment.WarningTMBistoxicavoiddirectcontactwithhands.Disposeoffproperly.Ifadarkbluecolourdevelopswithinafewminutesafterpreparation,thisindicatesthattheTMBsolutionhasbeencontaminatedandmustbediscarede.Readabsorbanceswithin1houraftercompletionoftheassay.?Whenpipettingreagents,maintainaconsistentorderofadditionfromwell-to-well.Thiswillensureequalincubationtimesforallwells.?Respectincubationtimesdescribedintheassayprocedure.SPECIMENCOLLECTION\PROCESSINGANDSTORAGE?Serum---Avoidanyinintentionalstimulationofthecellsbytheprocedure.Usepyrogen\endotoxinfreecollectingtubes.Serum首ldberemovedrapidlyandcarefullyfromtheredcellsafterclothing.Forthat,afterclothing,centrifugeatapproximately1000×gfor10minandremoveserum.?Plasma---EDTA\citrateandheparinplasmacanbeassayed.Spinsamplesat1000×gfor30minremoveparticulates.Harvestplasma.?Cellculturesupernatants---Removeparticulatesandaggregatesbyspinningatapproximately1000×gfor10min.?Storage---Ifnotanalyzedshortlyaftercollection,samples首ldbealiquoted(250-500ul)toavoidfreeze-thawcyclesandstoredfrozenat-70℃.Avoidmultiplefreeze-thawcyclesoffrozenspecimens.Whenpossible,avoiduseofbadlyhemolyzedorlipemicsera.Iflargeamountsofparticlesarepresent,this首ldberemovedpriortoassaybycentrifugationorfiltration.?Recommendation---Donotthawbyheatingat37℃or56℃.Thawatroomtemperatureandmakesurethatsampleiscompletelythawedandhomogenousbeforeassaying.PREPARATIONOFREAGENTS?Standards:Standardhavetobereconstituledwiththevolumeofstandardbufferdiluentindicatedonthevial.Thisreconstitutionproducesastocksolutionof400nmol/Lα-glucosidase.Allowstandardtostandfor5?minuteswithgentleswir领priortomakingdilutions.Serialdilutionsofstandardmustbemadebeforeeachassysandcannotbestored.400nmol/L（6Standard）Originaldensity50ul。200nmol/L（5Standard）100ul6Standard+100uldiludent100nmol/L（4Standard）100ul5Standard+100uldiludent50nmol/L（3Standard）100ul4Standard+100uldiludent25nmol/L（2Standard）100ul3Standard+100uldiludent12.5nmol/L（1Standard）100ul2Standard+100uldiludent0nmol/LBlankControl50ul。?Washingbuffer50×concentrate:Dilute50timesindistilledwater.ASSAYMETHOD?Beforeuse,mixallreagentsthoroughlywithoutmakingfoam.?Determinethenumberofmicrowellstripsrequiredtotestthedesirednumberofsamples,plusappropriatenumberofwellsneededforrunningblanksstandards.Eachsample,standardandblank首ldbeassayedinduplicate.Removesufficientmicrowellstripsfromthepouch.?Add50ulofstandarddiluenttostandardwellsB1,B2,C1,C2,D1,D2,E1,E2,F1,F2.Reconstitutestandardvialwiththeappropriatevolumeasdescribedinthechapterreagentspreparation.Preparation.Pipet100ulofstandardintowellsA1andA2(seeplateschemebelow).Transfer50ulfromA1andA2toB1andB2wells.Mixthecontentsbyrepeatedaspirationsandejections.Takecarenottoscratchtheinnersurfaceofmicrowells.RepatthisprocedurefromthewellsB1,B2towellsC1,C2andfromwellsC1,C2toD1,D2andsooncreatingtwoparallelrowsofα-glucosidasestandarddilutionsranging，Add50ulofstandarddiluenttotheblandwells.?Dilutesamples1:1distribing50ulofsampleinto50ulofdilluent,Add50ulofdilutedsampletowells..?Add50ulofdilutedbiotinylatedanti-α-glucosidasetoallwells.?Coverwithaplatevoverandincubatefor1hourat37℃.?Removethecoverandwashtheplateasfollows:⑴aspiratetheliquidfromeachwell,⑵dispensse0.3mlofwashingsolutionintoeachwell.⑶Aspirateagainthecontetofeachwellafter0.5minute.⑷Repeatsteps⑵and⑶threetimes.?Distribute60ulofstreptavidin-HRPsolutiontoallwells,includingblankwells.?Coverandincubate30minat37℃.?Removethecoverandemptywells,Washmicrowellstripsaccordingtostep,Proceedimmediatelytothenextstep.?Add50ulSubstrateAandSubstrateBtoeachwell。Incubatefor10minat37℃。?Theenzyme-substratereactionisstoppedbyquicklypipetting50ulofH2SO4.stopreagentintoeachwell,includingtheblankwells,tocompletelyanduniformlyinactivatetheenzyme.ResultsmustberedimmediatelyaftertheadditionofH2SO4.?Readabsorbanceofeachwellonaspectrophotometerusing450nmastheprimarywavelengthandoptionally620nm(610nmto650nmisacceptable)asthereferencewavelength.GoatAnti-Mouseα-glucosidaseSUGGESTEDPLATESCHEMEStandardconcentrations（nmol/L）A400400samplesamplesamplesamplesamplesamplesamplesamplesamplesampleB200200samplesamplesamplesamplesamplesamplesamplesamplesamplesampleC100100samplesamplesamplesamplesamplesamplesamplesamplesamplesampleD5050samplesamplesamplesamplesamplesamplesamplesamplesamplesampleE2525samplesamplesamplesamplesamplesamplesamplesamplesamplesampleF12.512.5samplesamplesamplesamplesamplesamplesamplesamplesamplesampleG00samplesamplesamplesamplesamplesamplesamplesamplesamplesampleHsamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesampleLIMITATIONSOFTHEPROCEDUREDonotextrapolatethestandardcurvebeyondthemaxstandardcurvepoint.Thedose-responseisnon-linearinthisregionandgoodaccruacyisdifficulttoobtain.CALCULATIONOFRESULTSTheminimumdetectableconcentrationinthisassayisestimatedtobe1.0nmol/L

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Goat Anti-Mouse α-glucosidase

GoatAnti-Mouseα-glucosidaseStorage:2-8°CPackagesize:96determinationsPRINCIPLEOFTHEMETHODTheα-glucosidasekitisasolidphasephasesandwichenzymelinkedimmunosorbentassay(ELISA).Samples,includingstandardsofknownα-glucosidaseconcentrationsandunknownsarepipettedintothesewells.Duringthefirstincubation,theα-glucosidaseantigenandabiotinylatedmonoclonalantibodyspecificforα-glucosidasearesimultaneouslyincubated.Afterwashing,theenzyme(streptavidin-peroxydase)isadded.Afterincubationandwashingtoremovealltheunboundenzyme,asubstratesolutionwhichisactingontheboundenzymeisaddedtoinduceacolouredreactionproduct.Theintensityofthiscolouredproductisdirectlyproportionaltotheconcentrationofα-glucosidasepresentinthesamples.REAGENTSPROVIDEDANDRECONSTTTUTIONREAGENTS（Storeat2-8℃）1×96WELLS0.5×96WELLSRECONSTTTUTION96/48-wellsmicrotiterplates10.5Ready-to-usePlastivcover21Ready-to-useStandard:400nmol/L1Vials(0.6ml)0.5Vials(0.3ml)Seereagentspreparationonpage3Blankcontrol1Vials(1.0ml)1Vials(0.5ml)Ready-to-useStandardDiluent1Vials(4.0ml)1Vials(2.0ml)Ready-to-useBiotinylatedanti-α-glucosidase1Vials(6.0ml)1Vials(3.0ml)Ready-to-useStreptavidin-HRP1Vials(8.0ml)1Vials(4.0ml)Ready-to-useWashingBuffer1Vials(20ml)1Vials(10ml)50×concentrateSubstrateA1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSubstrateB1Vials（6.0ml)1Vials(3.0ml)Ready-to-useStoppingSolution1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSampleDiluent1Vials(12ml)1Vials(6.0ml)Ready-to-useGoatAnti-Mouseα-glucosidaseMATERIALREQUIREDBUTNOTPROVIDED?Distilledwater?Pipettes:10ul、50ul、100ul、200ul、1000ul。?Vortexmixerandmagneticstirrer.SAFETY?Forresearchuseonly?AvoidanyskincontactwithH2SO4andTMB.Incaseofcontact,washthoroughlywater.?Donoteat,drink,smokeorapplycosmeticswherekitreagentsareused.?Donotpipettebymouth.PROCEDURALNOTES/LAB.QUALITYCONTROL?Whennotinuse,kitcomponents首ldbestoredrefrigeratedorfrozenasindicatedonvialsorbottles.Allreagents首ldbewarmedtoroomtemperaturebeforeuse.Lyophilizedstandards首ldbediscardedafteruse.?Oncethedesirednumberofstripshasbeenremoved,immediatelyresealthebagtoprotecttheremainingstripsfromedterioration.?Coverorcapallreagentswhennotinuse.?Donotmisorinterchangereagentsbetweendifferentlots.?Donotusereagentsbeyondtheexpirationdateofthekit.?Useacleandisposableplasticpipettetipforeachreagent,standard,orspecimenadditioninordertoavoidcross-contamination,forthedispensingofH2SO4andsubstratesolution,avoidpipetteswithmetalparts.?Useacleanplasticcontainertopreparethewashingsolution.?Thoroughlymixthereagentsandsamplesbeforeusebyagitationorswir领.?Allresidualwashingliquidmustbedrainedfromthewellsbyefficientaspirationorbydecantationfollowedbytappingtheplateforcefullyonabsorbentpaper.Neverinsertabsorbentpaperdirectlyintothewells.?TheTMBsolutionislightsensitive.Avoidprolongedexposuretolight,also,avoidcontactoftheTMBsolutionwithmetaltopreventcolourdevelopment.WarningTMBistoxicavoiddirectcontactwithhands.Disposeoffproperly.Ifadarkbluecolourdevelopswithinafewminutesafterpreparation,thisindicatesthattheTMBsolutionhasbeencontaminatedandmustbediscarede.Readabsorbanceswithin1houraftercompletionoftheassay.?Whenpipettingreagents,maintainaconsistentorderofadditionfromwell-to-well.Thiswillensureequalincubationtimesforallwells.?Respectincubationtimesdescribedintheassayprocedure.SPECIMENCOLLECTION\PROCESSINGANDSTORAGE?Serum---Avoidanyinintentionalstimulationofthecellsbytheprocedure.Usepyrogen\endotoxinfreecollectingtubes.Serum首ldberemovedrapidlyandcarefullyfromtheredcellsafterclothing.Forthat,afterclothing,centrifugeatapproximately1000×gfor10minandremoveserum.?Plasma---EDTA\citrateandheparinplasmacanbeassayed.Spinsamplesat1000×gfor30minremoveparticulates.Harvestplasma.?Cellculturesupernatants---Removeparticulatesandaggregatesbyspinningatapproximately1000×gfor10min.?Storage---Ifnotanalyzedshortlyaftercollection,samples首ldbealiquoted(250-500ul)toavoidfreeze-thawcyclesandstoredfrozenat-70℃.Avoidmultiplefreeze-thawcyclesoffrozenspecimens.Whenpossible,avoiduseofbadlyhemolyzedorlipemicsera.Iflargeamountsofparticlesarepresent,this首ldberemovedpriortoassaybycentrifugationorfiltration.?Recommendation---Donotthawbyheatingat37℃or56℃.Thawatroomtemperatureandmakesurethatsampleiscompletelythawedandhomogenousbeforeassaying.PREPARATIONOFREAGENTS?Standards:Standardhavetobereconstituledwiththevolumeofstandardbufferdiluentindicatedonthevial.Thisreconstitutionproducesastocksolutionof400nmol/Lα-glucosidase.Allowstandardtostandfor5?minuteswithgentleswir领priortomakingdilutions.Serialdilutionsofstandardmustbemadebeforeeachassysandcannotbestored.400nmol/L（6Standard）Originaldensity50ul。200nmol/L（5Standard）100ul6Standard+100uldiludent100nmol/L（4Standard）100ul5Standard+100uldiludent50nmol/L（3Standard）100ul4Standard+100uldiludent25nmol/L（2Standard）100ul3Standard+100uldiludent12.5nmol/L（1Standard）100ul2Standard+100uldiludent0nmol/LBlankControl50ul。?Washingbuffer50×concentrate:Dilute50timesindistilledwater.ASSAYMETHOD?Beforeuse,mixallreagentsthoroughlywithoutmakingfoam.?Determinethenumberofmicrowellstripsrequiredtotestthedesirednumberofsamples,plusappropriatenumberofwellsneededforrunningblanksstandards.Eachsample,standardandblank首ldbeassayedinduplicate.Removesufficientmicrowellstripsfromthepouch.?Add50ulofstandarddiluenttostandardwellsB1,B2,C1,C2,D1,D2,E1,E2,F1,F2.Reconstitutestandardvialwiththeappropriatevolumeasdescribedinthechapterreagentspreparation.Preparation.Pipet100ulofstandardintowellsA1andA2(seeplateschemebelow).Transfer50ulfromA1andA2toB1andB2wells.Mixthecontentsbyrepeatedaspirationsandejections.Takecarenottoscratchtheinnersurfaceofmicrowells.RepatthisprocedurefromthewellsB1,B2towellsC1,C2andfromwellsC1,C2toD1,D2andsooncreatingtwoparallelrowsofα-glucosidasestandarddilutionsranging，Add50ulofstandarddiluenttotheblandwells.?Dilutesamples1:1distribing50ulofsampleinto50ulofdilluent,Add50ulofdilutedsampletowells..?Add50ulofdilutedbiotinylatedanti-α-glucosidasetoallwells.?Coverwithaplatevoverandincubatefor1hourat37℃.?Removethecoverandwashtheplateasfollows:⑴aspiratetheliquidfromeachwell,⑵dispensse0.3mlofwashingsolutionintoeachwell.⑶Aspirateagainthecontetofeachwellafter0.5minute.⑷Repeatsteps⑵and⑶threetimes.?Distribute60ulofstreptavidin-HRPsolutiontoallwells,includingblankwells.?Coverandincubate30minat37℃.?Removethecoverandemptywells,Washmicrowellstripsaccordingtostep,Proceedimmediatelytothenextstep.?Add50ulSubstrateAandSubstrateBtoeachwell。Incubatefor10minat37℃。?Theenzyme-substratereactionisstoppedbyquicklypipetting50ulofH2SO4.stopreagentintoeachwell,includingtheblankwells,tocompletelyanduniformlyinactivatetheenzyme.ResultsmustberedimmediatelyaftertheadditionofH2SO4.?Readabsorbanceofeachwellonaspectrophotometerusing450nmastheprimarywavelengthandoptionally620nm(610nmto650nmisacceptable)asthereferencewavelength.GoatAnti-Mouseα-glucosidaseSUGGESTEDPLATESCHEMEStandardconcentrations（nmol/L）A400400samplesamplesamplesamplesamplesamplesamplesamplesamplesampleB200200samplesamplesamplesamplesamplesamplesamplesamplesamplesampleC100100samplesamplesamplesamplesamplesamplesamplesamplesamplesampleD5050samplesamplesamplesamplesamplesamplesamplesamplesamplesampleE2525samplesamplesamplesamplesamplesamplesamplesamplesamplesampleF12.512.5samplesamplesamplesamplesamplesamplesamplesamplesamplesampleG00samplesamplesamplesamplesamplesamplesamplesamplesamplesampleHsamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesampleLIMITATIONSOFTHEPROCEDUREDonotextrapolatethestandardcurvebeyondthemaxstandardcurvepoint.Thedose-responseisnon-linearinthisregionandgoodaccruacyisdifficulttoobtain.CALCULATIONOFRESULTSTheminimumdetectableconcentrationinthisassayisestimatedtobe1.0nmol/L[详细]
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Goat Anti-Pig Interleukin 6

GoatAnti-PigInterleukin6Storage:2-8°CPackagesize:96determinationsPRINCIPLEOFTHEMETHODTheIL-6kitisasolidphasephasesandwichenzymelinkedimmunosorbentassay(ELISA).Samples,includingstandardsofknownIL-6concentrationsandunknownsarepipettedintothesewells.Duringthefirstincubation,theIL-6antigenandabiotinylatedmonoclonalantibodyspecificforIL-6aresimultaneouslyincubated.Afterwashing,theenzyme(streptavidin-peroxydase)isadded.Afterincubationandwashingtoremovealltheunboundenzyme,asubstratesolutionwhichisactingontheboundenzymeisaddedtoinduceacolouredreactionproduct.TheintensityofthiscolouredproductisdirectlyproportionaltotheconcentrationofIL-6presentinthesamples.REAGENTSPROVIDEDANDRECONSTTTUTIONREAGENTS（Storeat2-8℃）1×96WELLS0.5×96WELLSRECONSTTTUTION96/48-wellsmicrotiterplates10.5Ready-to-usePlastivcover21Ready-to-useStandard:800pg/ml1Vials(0.6ml)0.5Vials(0.3ml)Seereagentspreparationonpage3Blankcontrol1Vials(1.0ml)1Vials(0.5ml)Ready-to-useStandardDiluent1Vials(4.0ml)1Vials(2.0ml)Ready-to-useBiotinylatedanti-IL-61Vials(6.0ml)1Vials(3.0ml)Ready-to-useStreptavidin-HRP1Vials(8.0ml)1Vials(4.0ml)Ready-to-useWashingBuffer1Vials(20ml)1Vials(10ml)50×concentrateSubstrateA1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSubstrateB1Vials（6.0ml)1Vials(3.0ml)Ready-to-useStoppingSolution1Vials(6.0ml)1Vials(3.0ml)Ready-to-useSampleDiluent1Vials(12ml)1Vials(6.0ml)Ready-to-useMATERIALREQUIREDBUTNOTPROVIDED?Distilledwater?Pipettes:10ul、50ul、100ul、200ul、1000ul。?Vortexmixerandmagneticstirrer.SAFETY?Forresearchuseonly?AvoidanyskincontactwithH2SO4andTMB.Incaseofcontact,washthoroughlywater.?Donoteat,drink,smokeorapplycosmeticswherekitreagentsareused.?Donotpipettebymouth.PROCEDURALNOTES/LAB.QUALITYCONTROL?Whennotinuse,kitcomponents首ldbestoredrefrigeratedorfrozenasindicatedonvialsorbottles.Allreagents首ldbewarmedtoroomtemperaturebeforeuse.Lyophilizedstandards首ldbediscardedafteruse.?Oncethedesirednumberofstripshasbeenremoved,immediatelyresealthebagtoprotecttheremainingstripsfromedterioration.?Coverorcapallreagentswhennotinuse.?Donotmisorinterchangereagentsbetweendifferentlots.?Donotusereagentsbeyondtheexpirationdateofthekit.?Useacleandisposableplasticpipettetipforeachreagent,standard,orspecimenadditioninordertoavoidcross-contamination,forthedispensingofH2SO4andsubstratesolution,avoidpipetteswithmetalparts.?Useacleanplasticcontainertopreparethewashingsolution.?Thoroughlymixthereagentsandsamplesbeforeusebyagitationorswir领.?Allresidualwashingliquidmustbedrainedfromthewellsbyefficientaspirationorbydecantationfollowedbytappingtheplateforcefullyonabsorbentpaper.Neverinsertabsorbentpaperdirectlyintothewells.?TheTMBsolutionislightsensitive.Avoidprolongedexposuretolight,also,avoidcontactoftheTMBsolutionwithmetaltopreventcolourdevelopment.WarningTMBistoxicavoiddirectcontactwithhands.Disposeoffproperly.Ifadarkbluecolourdevelopswithinafewminutesafterpreparation,thisindicatesthattheTMBsolutionhasbeencontaminatedandmustbediscarede.Readabsorbanceswithin1houraftercompletionoftheassay.?Whenpipettingreagents,maintainaconsistentorderofadditionfromwell-to-well.Thiswillensureequalincubationtimesforallwells.?Respectincubationtimesdescribedintheassayprocedure.SPECIMENCOLLECTION\PROCESSINGANDSTORAGE?Serum---Avoidanyinintentionalstimulationofthecellsbytheprocedure.Usepyrogen\endotoxinfreecollectingtubes.Serum首ldberemovedrapidlyandcarefullyfromtheredcellsafterclothing.Forthat,afterclothing,centrifugeatapproximately1000×gfor10minandremoveserum.?Plasma---EDTA\citrateandheparinplasmacanbeassayed.Spinsamplesat1000×gfor30minremoveparticulates.Harvestplasma.?Cellculturesupernatants---Removeparticulatesandaggregatesbyspinningatapproximately1000×gfor10min.?Storage---Ifnotanalyzedshortlyaftercollection,samples首ldbealiquoted(250-500ul)toavoidfreeze-thawcyclesandstoredfrozenat-70℃.Avoidmultiplefreeze-thawcyclesoffrozenspecimens.Whenpossible,avoiduseofbadlyhemolyzedorlipemicsera.Iflargeamountsofparticlesarepresent,this首ldberemovedpriortoassaybycentrifugationorfiltration.?Recommendation---Donotthawbyheatingat37℃or56℃.Thawatroomtemperatureandmakesurethatsampleiscompletelythawedandhomogenousbeforeassaying.PREPARATIONOFREAGENTS?Standards:Standardhavetobereconstituledwiththevolumeofstandardbufferdiluentindicatedonthevial.Thisreconstitutionproducesastocksolutionof800pg/mlIL-6.Allowstandardtostandfor5?minuteswithgentleswir领priortomakingdilutions.Serialdilutionsofstandardmustbemadebeforeeachassysandcannotbestored.800pg/ml（6Standard）Originaldensity50ul。400pg/ml（5Standard）100ul6Standard+100uldiludent200pg/ml（4Standard）100ul5Standard+100uldiludent100pg/ml（3Standard）100ul4Standard+100uldiludent50pg/ml（2Standard）100ul3Standard+100uldiludent25pg/ml（1Standard）100ul2Standard+100uldiludent0pg/mlBlankControl50ul。?Washingbuffer50×concentrate:Dilute50timesindistilledwater.ASSAYMETHOD?Beforeuse,mixallreagentsthoroughlywithoutmakingfoam.?Determinethenumberofmicrowellstripsrequiredtotestthedesirednumberofsamples,plusappropriatenumberofwellsneededforrunningblanksstandards.Eachsample,standardandblank首ldbeassayedinduplicate.Removesufficientmicrowellstripsfromthepouch.?Add50ulofstandarddiluenttostandardwellsB1,B2,C1,C2,D1,D2,E1,E2,F1,F2.Reconstitutestandardvialwiththeappropriatevolumeasdescribedinthechapterreagentspreparation.Preparation.Pipet100ulofstandardintowellsA1andA2(seeplateschemebelow).Transfer50ulfromA1andA2toB1andB2wells.Mixthecontentsbyrepeatedaspirationsandejections.Takecarenottoscratchtheinnersurfaceofmicrowells.RepatthisprocedurefromthewellsB1,B2towellsC1,C2andfromwellsC1,C2toD1,D2andsooncreatingtwoparallelrowsofIL-6standarddilutionsranging，Add50ulofstandarddiluenttotheblandwells.?Dilutesamples1:1distribing50ulofsampleinto50ulofdilluent,Add50ulofdilutedsampletowells..?Add50ulofdilutedbiotinylatedanti-IL-6toallwells.?Coverwithaplatevoverandincubatefor1hourat37℃.?Removethecoverandwashtheplateasfollows:⑴aspiratetheliquidfromeachwell,⑵dispensse0.3mlofwashingsolutionintoeachwell.⑶Aspirateagainthecontetofeachwellafter0.5minute.⑷Repeatsteps⑵and⑶threetimes.?Distribute60ulofstreptavidin-HRPsolutiontoallwells,includingblankwells.?Coverandincubate30minat37℃.?Removethecoverandemptywells,Washmicrowellstripsaccordingtostep,Proceedimmediatelytothenextstep.?Add50ulSubstrateAandSubstrateBtoeachwell。Incubatefor10minat37℃。?Theenzyme-substratereactionisstoppedbyquicklypipetting50ulofH2SO4.stopreagentintoeachwell,includingtheblankwells,tocompletelyanduniformlyinactivatetheenzyme.ResultsmustberedimmediatelyaftertheadditionofH2SO4.?Readabsorbanceofeachwellonaspectrophotometerusing450nmastheprimarywavelengthandoptionally620nm(610nmto650nmisacceptable)asthereferencewavelength.SUGGESTEDPLATESCHEMEStandardconcentrations（pg/ml）A800800samplesamplesamplesamplesamplesamplesamplesamplesamplesampleB400400samplesamplesamplesamplesamplesamplesamplesamplesamplesampleC200200samplesamplesamplesamplesamplesamplesamplesamplesamplesampleD100100samplesamplesamplesamplesamplesamplesamplesamplesamplesampleE5050samplesamplesamplesamplesamplesamplesamplesamplesamplesampleF2525samplesamplesamplesamplesamplesamplesamplesamplesamplesampleG00samplesamplesamplesamplesamplesamplesamplesamplesamplesampleHsamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesamplesampleLIMITATIONSOFTHEPROCEDUREDonotextrapolatethestandardcurvebeyondthemaxstandardcurvepoint.Thedose-responseisnon-linearinthisregionandgoodaccruacyisdifficulttoobtain.CALCULATIONOFRESULTSTheminimumdetectableconcentrationinthisassayisestimatedtobe1.0pg/ml[详细]
2018-09-27 10:00
产品样册
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Goat interleukin 17(IL-17)

Goat interleukin 17(IL-17)[详细]
2024-09-28 01:06
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Goat Anti-Rabbit Motilin Receptor

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2024-09-28 10:33
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人α葡萄糖苷酶（α-glucosidase）的注意事项

人α葡萄糖苷酶（α-glucosidase）的注意事项[详细]
2015-05-18 00:00
课件
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人α葡萄糖苷酶（α-glucosidase）的注意事项

人α葡萄糖苷酶（α-glucosidase）的注意事项[详细]
2015-03-25 00:00
专利
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人α葡萄糖苷酶（α-glucosidase）试剂盒使用方法

检测范围：96T7U/L-200U/L使用目的：本试剂盒用于测定人血清、血浆及相关液体样本中α葡萄糖苷酶（α-glucosidase）含量。实验原理本试剂盒应用双抗体夹心法测定标本中人α葡萄糖苷酶（α-glucosidase）水平。用纯化的人α葡萄糖苷酶（α-glucosidase）抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入α葡萄糖苷酶（α-glucosidase），再与HRP标记的α葡萄糖苷酶（α-glucosidase）抗体结合，形成抗体-抗原-酶标抗体复合物，经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色，并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的α葡萄糖苷酶（α-glucosidase）呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），通过标准曲线计算样品中人α葡萄糖苷酶（α-glucosidase）浓度。[详细]
2018-10-01 10:00
产品样册
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Goat Anti-Dog Tumor necrosis factor

Goat Anti-Dog Tumor necrosis factor [详细]
2014-04-22 00:00
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Goat anti- Guinea Pig Interleukin-2

Goat anti- Guinea Pig Interleukin-2[详细]
2014-04-21 00:00
产品样册
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Goat anti- Rabbit Inhibin B

Goat anti- Rabbit Inhibin B[详细]
2024-09-22 19:49
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Goat anti- Rabbit Nitric oxide

Goat anti- Rabbit Nitric oxide[详细]
2024-09-28 00:20
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Rat Anti- Goat Brucella Antibody

Rat Anti- Goat Brucella Antibody[详细]
2024-09-20 03:03
报价单
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APC anti-mouse CD16/32 - blocks Fc binding

eBioscience公司位于美国的亚哥，其产品被全世界实验室所采用。eBioscience产品包括人细胞表面抗体细胞信号，细胞因子；小鼠细胞表面抗原，细胞信号，细胞因子和大鼠细胞表面抗原，细胞因子；品种多，规格全，能满足各类实验需求。[详细]
2018-09-18 10:00
产品样册
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小鼠葡糖苷酶(-glucosidase)ELISA试剂盒

小鼠葡糖苷酶(-glucosidase)ELISA试剂盒[详细]
2013-12-09 00:00
实验操作
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人葡糖苷酶(-glucosidase)ELISA试剂盒

人葡糖苷酶(-glucosidase)ELISA试剂盒[详细]
2013-12-06 00:00
应用文章
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小鼠（Mouse） α-葡萄糖苷酶（α-glucosidase） ELISA 检测试剂盒说明书

小鼠(Mouse)α-葡萄糖苷酶(α-glucosidase)ELISA检测试剂盒本试剂仅供研究使用试验原理：α-glucosidase试剂盒是固相夹心法酶联免疫吸附实验（ELISA）.已知α-glucosidase浓度的标准品、未知浓度的样品加入微孔酶标板内进行检测。先将α-glucosidase和生物素标记的抗体同时温育。洗涤后，加入亲和素标记过的HRP。再经过温育和洗涤，去除未结合的酶结合物，然后加入底物A、B，和酶结合物同时作用。产生颜色。颜色的深浅和样品中α-glucosidase的浓度呈比例关系。试剂盒内容及其配制试剂盒成份（2-8℃保存）96孔配置48孔配置配制96/48人份酶标板1块板（96T）半块板（48T）即用型塑料膜板盖1块半块即用型标准品：400nmol/L1瓶（0.6ml）1瓶（0.3ml）按说明书进行稀稀空白对照1瓶（1.0ml）1瓶（0.5ml）即用型标准品稀释缓冲液1瓶（4.0ml）1瓶（2.0ml）即用型生物素标记的抗α-glucosidase抗体1瓶（6.0ml）1瓶（3.0ml）即用型亲和链酶素-HRP1瓶（8.0ml）1瓶（4.0ml）即用型洗涤缓冲液1瓶（20ml）1瓶（10ml）按说明书进行稀释底物A1瓶（6.0ml）1瓶（3.0ml）即用型底物B1瓶（6.0ml）1瓶（3.0ml）即用型终止液1瓶（6.0ml）1瓶（3.0ml）即用型标本稀释液1瓶（12ml）1瓶（6.0ml）即用型自备材料蒸馏水。加样器：5ul、10ul、50ul、100ul、200ul、500ul、1000ul。振荡器及磁力搅拌器等。安全性避免直接接触终止液和底物A、B。一旦接触到这些液体，请尽快用水冲洗。实验中不要吃喝、抽烟或使用化妆品。不要用嘴吸取试剂盒里的任何成份。操作注意事项试剂应按标签说明书储存，使用前恢复到室温。稀稀过后的标准品应丢弃，不可保存。实验中不用的板条应立即放回包装袋中，密封保存，以免变质。不用的其它试剂应包装好或盖好。不同批号的试剂不要混用。保质前使用。使用一次性的吸头以免交叉污染，吸取终止液和底物A、B液时，避免使用带金属部分的加样器。使用干净的塑料容器配置洗涤液。使用前充分混匀试剂盒里的各种成份及样品。洗涤酶标板时应充分拍干，不要将吸水纸直接放入酶标反应孔中吸水。底物A应挥发，避免长时间打开盖子。底物B对光敏感，避免长时间暴露于光下。避免用手接触，有毒。实验完成后应立即读取OD值。加入试剂的顺序应一致，以保证所有反应板孔温育的时间一样。按照说明书中标明的时间、加液的量及顺序进行温育操作。样品收集、处理及保存方法血清-----操作过程中避免任何细胞刺激。使用不含热原和内毒素的试管。收集血液后，1000×g离心10分钟将血清和红细胞迅速小心地分离。血浆-----EDTA、柠檬酸盐、肝素血浆可用于检测。1000×g离心30分钟去除颗粒。细胞上清液---1000×g离心10分钟去除颗粒和聚合物。组织匀浆-----将组织加入适量生理盐水捣碎。1000×g离心10分钟，取上清液保存------如果样品不立即使用，应将其分成小部分-70℃保存，避免反复冷冻。尽可能的不要使用溶血或高血脂血。如果血清中大量颗粒，检测前先离心或过滤。不要在37℃或更高的温度加热解冻。应在室温下解冻并确保样品均匀地充分解冻。试剂的准备标准品：标准品的系列稀释应在实验时准备，不能储存。稀释前将标准品振荡混匀。稀释比例按下表中进行：400nmol/L（6号标准品）原倍浓度不用稀释直接加入50ul。200nmol/L（5号标准品）100ul的原倍标准品加入100ul的标准品稀释液100nmol/L（4号标准品）100ul的5号标准品加入100ul的标准品稀释液50nmol/L（3号标准品）100ul的4号标准品加入100ul的标准品稀释液25nmol/L（2号标准品）100ul的3号标准品加入100ul的标准品稀释液12.5nmol/L（1号标准品）100ul的2号标准品加入100ul的标准品稀释液0nmol/L（空白对照）原始浓度不用稀释直接加入50ul。洗涤缓冲液（50×）的稀释：蒸馏水50倍稀释。操作步骤使用前，将所有试剂充分混匀。不要使液体产生大量的泡沫，以免加样时加入大量的气泡，产生加样上的误差。根据待测样品数量加上标准品的数量决定所需的板条数。每个标准品和空白孔建议做复孔。每个样品根据自己的数量来定，能使用复孔的尽量做复孔。标本用标本稀释液1：1稀释后加入50ul于反应孔内。加入稀释好后的标准品50ul于反应孔、加入待测样品50ul于反应孔内。立即加入50ul的生物素标记的抗体。盖上膜板，轻轻振荡混匀，37℃温育1小时。甩去孔内液体，每孔加满洗涤液，振荡30秒，甩去洗涤液，用吸水纸拍干。重复此操作3次。如果用洗板机洗涤，洗涤次数增加一次。每孔加入60ul的亲和链酶素-HRP，轻轻振荡混匀，37℃温育30分钟。甩去孔内液体，每孔加满洗涤液，振荡30秒，甩去洗涤液，用吸水纸拍干。重复此操作3次。如果用洗板机洗涤，洗涤次数增加一次。每孔加入底物A、B各50ul，轻轻振荡混匀，37℃温育10分钟。避免光照。取出酶标板，迅速加入50ul终止液，加入终止液后应立即测定结果。在450nm波长处测定各孔的OD值。建议使用的实验方案标准品浓度（nmol/L）A400400样品样品样品样品样品样品样品样品样品样品B200200样品样品样品样品样品样品样品样品样品样品C100100样品样品样品样品样品样品样品样品样品样品D5050样品样品样品样品样品样品样品样品样品样品E2525样品样品样品样品样品样品样品样品样品样品F12.512.5样品样品样品样品样品样品样品样品样品样品G00样品样品样品样品样品样品样品样品样品样品H样品样品样品样品样品样品样品样品样品样品样品样品局限6号标准品以上的结果为非线性的，根据此标准曲线无法得到极ng确的结果。试剂盒性能1.灵敏度：Z小的检测浓度小于1号标准品。稀释度的线性。样品线性回归与预期浓度相关系数R值为0.990。2.特异性：不与其它细胞因子反应。3.重复性：板内、板间变异系数均小于10%。结果判断与分析1、仪器值：于波长450nm的酶标仪上读取各孔的OD值2、以吸光度OD值为纵坐标（Y），相应的α-glucosidase标准品浓度为横坐标（X），做得相应的曲线，样品的α-glucosidase含量可根据其OD值由标准曲线换算出相应的浓度。3、检测值范围：0-400nmol/L4、敏感度：1.0nmol/L[详细]
2018-09-14 10:00
产品样册
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该会员其他资料: 如何选择抗体--抗体选择指南; 人软骨糖蛋白39（HC gp-39）ELISA试剂盒说明书; 薄荷脑说明书; 新方法可快速廉价诊断肺结核; 菸酸缺乏病的发病机理介绍; 人β淀粉样蛋白1-42（Aβ1-42）Elisa试剂盒注意事项说明; 抗体催化的化学反应及制备; 关于水杨酸的美容效果介绍; 甲氨蝶呤的作用介绍; 抗体酶

Goat Anti-Mouse α-glucosidase

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