Description:
Transcriptionfactors(TFs)areagroupofcellularproteinsthatplayessentialrolesinregulatinggeneexpression.Often,aspecificcellularsignalpathwaycanactivatemultipleTFsandtheexpressionofaspecificgeneisundercontrolofmultipleTFs.Therefore,monitoringtheactivationofmultipleTFssimultaneouslyisveryimportant. DifferentfromTFActivationProfilingPlateArrayI,Signosis’TFactivationprofilingplatearrayIIisusedformonitoring96differentTFsincludingNFkB,AP1,HIF1,andp53.Two96-wellplatesareprovidedforthecomparisonoftwosamples.ApplicableGrid:
ListofApplicableTFs:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | AP1 | CDP | GATA | NF-1 | Pit | Stat3 | XBP | FOXG1 | HoxA-5 | NRF2(A) | ProX1 | SOX2 |
| B | AP2 | CREB | GR/PR | NFAT | PPAR | Stat4 | AP3 | FOXO1 | HSF | Oct-1 | RB | SOX9 |
| C | AR | E2F-1 | HIF | NF-E2 | PXR | Stat5 | AP4 | FREAC-2 | KLF4 | Pax2 | RUNX | SOX18 |
| D | ATF2 | EGR | HNF4 | NFkB | SMAD | Stat6 | COUP-TF | Gli-1 | MyoD | Pax3 | ROR(RZR) | SRY |
| E | Brn-3 | ER | IRF | OCT4 | Sp1 | TCF/LEF | ELK | Gfi-1 | MZF | Pax8 | RXR | TFE3 |
| F | CEBP | Ets | MEF2 | p53 | SRF | YY1 | FOXA1 | HEN(NSCL-1) | Nkx2-5 | PIT1 | SF-1 | USF-1 |
| G | CAR | FAST-1 | Myb | Pax-5 | SATB1 | TR | FOXC1 | HNF-1 | Nkx3-2 | PLAG1 | SMUC | VDR |
| H | CBF | GAS/ISRE | Myc-Max | Pbx1 | Stat1 | TFIID | FOXD3 | HOX4C | NRF1 | MEF1 | Snail | WT1 |
Principle:
Signosis’TFActivationProfilingArraysareusedformonitoringtheactivitiesofmultipleTFssimultaneously.Inthetechnology,aseriesofbiotin-labeledprobesaremadebasedontheconsensussequencesofTFDNA-bindingsites.Whentheprobemixturesincubatewithnuclearextracts,individualprobeswillfindtheircorrespondingTFsandformTF/probecomplexes,whichcanbeeasilyseparatedfromfreeprobesthroughasimplespincolumnpurification.Theboundprobesarethendetachedfromthecomplexesandanalyzedthroughhybridizationwithaplate,whicheachwellisspecificallypre-coatedwithcomplementarysequencesoftheprobes.
Data:
TFactivationprofilingplatearrayIIanalysistheactivitiesof96TFs.HeLacellsweretreatedwithandwithoutPMA.ThenuclearextractswerepreparedandsubjectedtoTFactivationprofilingplatearray.
