In Vitro Budding and Floatation based enrichment of Mitochondria-derived Vesicles for Proteomics from Rat Heart
Mitochondria-derived vesicles (MDVs) are a novel class of vesicles whose biogenesis and trafficking are poorly characterized. Here, we describe the protocol to in vitro generate MDVs from mitochondria isolated from animal heart tissues. The protocol can be coupled with Transmission Electron Microscopy (TEM), proteomics or western blotting as final readouts. The protocol is optimized to improve the yield and purity of vesicles for downstream proteomics and facilitates selection of key regulators to be validated within cells. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=196 SRC="FIGDIR/small/583262v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@1095430org.highwire.dtl.DTLVardef@1763db2org.highwire.dtl.DTLVardef@1cc7e2dorg.highwire.dtl.DTLVardef@6fcdb0_HPS_FORMAT_FIGEXP M_FIG C_FIG The protocol has been adapted and further modified from (Soubannier et al., 2012).