The Mitochondria Isolation Kit, mouse tissue facilitates the isolation of functional and viable mitochondria from mouse tissue. The isolation protocol is based on the renowned MACS Technology, which enables fast isolation of high purity and high yield mitochondria.
Details
Detailed procedure
Tissues can be homogenized with the gentleMACS or gentleMACS Octo Dissociator in combination with the Mitochondria Extraction Kit – Tissue. After one-step homogenization of the tissue and lysis of the cell suspension, mitochondria are magnetically labeled with Anti-TOM22 MicroBeads, mouse, which bind to the translocase of the outer mitochondrial membrane 22 protein (TOM22). The sample is loaded onto an LS Column placed in a MidiMACS or QuadroMACS Separator. After washing, only the magnetically labeled mitochondria are retained on the column. The column is removed from the separator and functional mouse mitochondria are eluted from the column.
Downstream applications
After isolation mitochondria can be further subjected to Western blot analysis1,6 or downstream functional assays, including measurement of O2, membrane potential, and ATP4-5,7, or respiratory control4. Growing evidence also suggests that isolated mitochondria are well suited for mitochondrial RNA expression profiling studies2,3.
