Materials:2xassaybuffer,200µMmethylatedpeptidesubstrate(BPScatalognumber50101),LSD1(BPScatalognumber50100),AmplexRedreagent,peroxidase
Step1:
1)Addtheentirereactionmixturetoa96wellplate:20µlLSD1(10-15ng/µl)5µlof200µMmethylatedpeptidesubstrate25µlof2xLSD1assaybuffer
2)Incubatefor15-30minatroomtemperature.
Step2:
1)Immediatelybeforeuse:add50µlof10mMAmplexRedstockand100µlof10U/mlperoxidaseto4.85mlof1xLSD1buffer,mix.
2)Add50µlofmixturetoeachwell.Mixwell.
3)Readfluorescenceatex=530nmandem=590nm.
12monthsfromdateofreceipt,whenstoredasdirected
ApplicationReference(s):
1. 4SC-202asaPotentialTreatmentforthePediatricBrainTumorMedulloblastoma(2017)
2. CrystalstructureofthehistonelysinespecificdemethylaseLSD1complexedwithtetrahydrofolate(2014)
Important: UseofthissubstraterequiresPeroxidaseRxnGradeLSD1, iforderingLSD1fromBPS,pleaseincludearequestfor"PeroxidaseRxnGradeLSD1"withyourorder.
