Components:- MitochondriaIsolationKit,mousetissue(130-096-946)
- MidiMACSSeparator(130-042-302)
- MACSMultiStand(130-042-303)
Overview
TheMitochondriaIsolationKit,mousetissuefacilitatestheisolationoffunctionalandviablemitochondriafrommousetissue.TheisolationprotocolisbasedontherenownedMACSTechnology,whichenablesfastisolationofhighpurityandhighyieldmitochondria.
Details
Detailedprocedure
TissuescanbehomogenizedwiththegentleMACSorgentleMACSOctoDissociatorincombinationwiththeMitochondriaExtractionKit–Tissue.Afterone-stephomogenizationofthetissueandlysisofthecells
USPension,mitochondriaaremagneticallylabeledwithAnti-TOM22MicroBeads,mouse,whichbindtothetranslocaseoftheoutermitochondrialmembrane22protein(TOM22).ThesampleisloadedontoanLSColumnplacedinaMidiMACSorQuadroMACSSeparator.Afterwashing,onlythemagneticallylabeledmitochondriaareretainedonthecolumn.Thecolumnisremovedfromtheseparatorandfunctionalmousemitochondriaareelutedfromthecolumn.
Downstreamapplications
AfterisolationmitochondriacanbefurthersubjectedtoWesternblotanalysis1,6ordownstreamfunctionalassays,includingmeasurementofO2,membranepotential,andATP4-5,7,orrespiratorycontrol4.GrowingevidencealsosuggeststhatisolatedmitochondriaarewellsuitedformitochondrialRNAexpressionprofilingstudies2,3.