| InputType: | UnisolatedSamples |
| ResearchArea: | Chromatin&Transcription |
| TargetApplication: | SampleIsolation |
| VesselFormat: | Columns/Tubes |
| 100%Guarantee: | 6months |
The ChromaFlash™ChromatinExtractionKit isacompletesetofoptimizedbuffersandreagentsforisolatingchromatinorDNA-proteincomplexfrommammaliancellsortissuesinasimpleandrapidformat.Chromatinpreparedbythiskitcanbeusedinavarietyofchromatinimmunoprecipitationmethods.ItisalsotherecommendedmethodforobtainingchromatinrequiredbyEpigentek’sone-hourChIPmethodusingthe ChromaFlash™One-StepChIPKit.Theisolatedchromatincanalsobeusedinotherchromatin-relatedapplicationssuchasinvitroprotein-DNAbindingassaysandnuclearenzymeassays.
- Extremelyfastprocedure:theentireprocedurefromcell/tissuesampletoready-to-usechromatinislessthan60minutes.
- ConvenientandflexIBLe:thekitissuitableforpreparingbothnativechromatinandcross-linkedchromatinfrommonolayerorsUSPensioncells,orfromtissues.
- Unshearedchromatinmakesitcustomizableforvariousanalysisworkflowsthatrequireeitherintactorfragmentedchromatin,includingChIP,invitroprotein-DNAinteractionanalysis,nuclearenzymeassay,etc.
BackgroundInformation
Chromatinimmunoprecipitaton(ChIP)offersanadvantageoustoolforstudyingprotein-DNAinteraction.WithChIP,theexperimentercandetermineifaspecificproteinbindstothespecificsequencesofageneinlivingcellsbycombiningwithPCR(ChIP-PCR),microarray(ChIP-chip),orsequencing(ChIP-Seq)techniques.Forexample,themeasurementoftheamountofmethylatedhistoneH3atlysine9(meH3-K9)associatedwithaspecificgenepromoterregionundervariousconditionscanbeachievedthroughaChIP-PCRassay,whilerecruitmentofmeH3-K9tothepromotersonagenome-widescalecanbedetectedbyChIP-chip.Inparticular,theChIPmethodwithspecificantibodiesdirectlyagainstvarioustranscriptionalfactorsiswidelydemanded.
Principle&Procedure
TheChromaFlash™ChromatinExtractionKitcontainsallreagentsrequiredforcarryingoutsuccessfulchromatinextractiondirectlyfrommammaliancellsortissues.Cellmembranesofthesample,withorwithoutcross-linking,arebrokendownusingtheprovidedlysisbuffer.ChromatinorDNA-proteincomplexisthenextractedwiththeextractionbuffer.Theextractedchromatincanthenbedilutedwithchromatinbufferandstoredattheappropriatetemperature.
StartingMaterials&InputAmount
Startingmaterialscanincludevarioustissueorcellsamplessuchascellsfromflaskormicroplateculturedcells,freshandfrozentissues,etc.Theamountofcellsandtissuesforeachpreparationcanbe1x105 to5x106 cellsand10mgto200mg,respectively.Foroptimalpreparation,theinputamountshouldbe1to5x106 cellsor50to200mgtissues. Yieldofchromatinisapproximately4ugper106 cellsorper50mgtissues.
10XLysisBuffer
ExtractionBuffer
ChromatinBuffer
ProteaseInhibitorCocktails(1000X)
UserGuide
