Rehydratewww
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www.
ptglab.
comPRODUCT-SPECIFICPROTOCOLSIHC(10782-2-AP)SampletypeSamplepreparationAntigenRetrievalBlockingbufferPrimaryantibodydilutionIncubationtimeSignaldetectionmousebraintissueParaffin-embeddedTris-EDTAbuffer(pH9)5%goatseruminTBS1:20001.
5hatroomtempDako/AgilentEnvisionkitmousebraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitmousebraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitratbraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitPROTOCOL(moredetailsavailableuponrequest):1.
Deparaffinizesectionsin2bathsofxylenefor10mineach.
2.
Rehydratesectionsbysequentialincubationwith100%,95%,80%,and60%ethanol,5minforeachbath.
3.
Rinsesectionswithdistilledwater3timesfor3mineach.
AntigenRetrieval(checktableabove)4.
Transfersectionstoacontainerandcoverwithantigenretrievalsolutionaccordingtothetableabove.
5.
Heatslidesinamicrowaveonmediumpowerfor10min.
6.
Allowslidestocoolinthebufferfor35min.
IncubationwithPrimaryAntibodyandSignalDetection7.
Rinseslides3timeswith1XTBSfor3mineach.
8.
Incubateslideswith3%H2O2solution(dilutedindistilledwater)for10mintoquenchendogenousperoxidaseactivity.
9.
Rinseslides3timeswith1XTBSfor3mineach.
10.
Blockthesectionsatroomtemperaturefor1hinBlockingbuffer.
11.
Incubatesectionswithprimaryantibodyin1XTBS12.
Rinseslides3timeswith1XTBSfor3mineach.
13.
Performsignaldetectionprotocolaccordingtothemanufacturer'sinstructions.
14.
Immerseslidesinabathofhematoxylintostainthenucleifor3min.
15.
Rinseslidesgentlyinadistilledwaterbath.
16.
Transferslidestoasolutioncontaining1%HCland99%ethanolfor10sec,andthenintodistilledwaterimmediately.
17.
Immerseslidessequentiallyinto60%,80%,90%,and100%ethanolbathfor5mineach.
18.
Incubateslidesinxylenebathfor5min,remove,thenincubateagainfor5min.
19.
Mountthesectionwithasmalldropofneutralbalsamandaddacoverslip.
Air-dryslidesinthehood.
20.
Examineslidesunderamicroscope.
Visitptglab.
comforfullproductrange
ptglab.
comPRODUCT-SPECIFICPROTOCOLSIHC(10782-2-AP)SampletypeSamplepreparationAntigenRetrievalBlockingbufferPrimaryantibodydilutionIncubationtimeSignaldetectionmousebraintissueParaffin-embeddedTris-EDTAbuffer(pH9)5%goatseruminTBS1:20001.
5hatroomtempDako/AgilentEnvisionkitmousebraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitmousebraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitratbraintissueParaffin-embeddedNone5%goatseruminTBS1:2001.
5hatroomtempDako/AgilentEnvisionkitPROTOCOL(moredetailsavailableuponrequest):1.
Deparaffinizesectionsin2bathsofxylenefor10mineach.
2.
Rehydratesectionsbysequentialincubationwith100%,95%,80%,and60%ethanol,5minforeachbath.
3.
Rinsesectionswithdistilledwater3timesfor3mineach.
AntigenRetrieval(checktableabove)4.
Transfersectionstoacontainerandcoverwithantigenretrievalsolutionaccordingtothetableabove.
5.
Heatslidesinamicrowaveonmediumpowerfor10min.
6.
Allowslidestocoolinthebufferfor35min.
IncubationwithPrimaryAntibodyandSignalDetection7.
Rinseslides3timeswith1XTBSfor3mineach.
8.
Incubateslideswith3%H2O2solution(dilutedindistilledwater)for10mintoquenchendogenousperoxidaseactivity.
9.
Rinseslides3timeswith1XTBSfor3mineach.
10.
Blockthesectionsatroomtemperaturefor1hinBlockingbuffer.
11.
Incubatesectionswithprimaryantibodyin1XTBS12.
Rinseslides3timeswith1XTBSfor3mineach.
13.
Performsignaldetectionprotocolaccordingtothemanufacturer'sinstructions.
14.
Immerseslidesinabathofhematoxylintostainthenucleifor3min.
15.
Rinseslidesgentlyinadistilledwaterbath.
16.
Transferslidestoasolutioncontaining1%HCland99%ethanolfor10sec,andthenintodistilledwaterimmediately.
17.
Immerseslidessequentiallyinto60%,80%,90%,and100%ethanolbathfor5mineach.
18.
Incubateslidesinxylenebathfor5min,remove,thenincubateagainfor5min.
19.
Mountthesectionwithasmalldropofneutralbalsamandaddacoverslip.
Air-dryslidesinthehood.
20.
Examineslidesunderamicroscope.
Visitptglab.
comforfullproductrange
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