Method:
____1.DryMembraneaftertransfer.Rehydratenitrocellulosemembranefor~5minutesin
PBS/TBSorifusingPVDFmembrane,brie?yinmethanol,thenPBS/TBSfor5minutes.
____2.Incubate1houratroomtemperatureinblocker(nodetergent)withgentleagitation.
On@___________________
O?@___________________
____3.Decantblockandaddprimaryantibodydiluted1:_______inblocker+0.2%Tween?20.
Incubate1houratroomtemperaturewithgentleagitation.
On@___________________
O?@___________________
____4.Decantandwash4X,5minuteseachwash,withagitation,using1XPBS/TBS+0.1%
Tween20
qWashstep1qWashstep2qWashstep3qWashstep4
____5.Decant?nalwashandaddsecondaryantibodydiluted1:_________inblocker+0.2%
Tween20.Incubate1houratroomtemperatureinthedarkwithgentleagitation.
On@___________________
O?@___________________
____6.Decantandwash4X,5minuteseachwash,withagitation,using1XPBS/TBS+0.1%
Tween20
qWashstep1qWashstep2qWashstep3qWashstep4
____7.Finalwashin1XPBS/TBSonly,5minutesatroomtemperaturewithgentleagitation.
qFinalwashstep
____8.Addsubstrate:_____________mLfor____________minutes
____9.Imageon__________________________________Parameters:___________________________
Notes:_______________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
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