Isolate mitochondrial proteins from soft and hard tissue.
The Thermo Scientific Mitochondria Isolation Kit for Tissue enables isolation of intact mitochondria from soft and hard tissue samples in about an hour using reagent-based or Dounce homogenization methods.
This kit provides for two methods for mitochondria isolation. The first method uses a unique reagent-based procedure that enables simultaneous multi-sample processing. The second method relies on traditional Dounce homogenization for tissue disruption and subsequent isolation of the organelle. Both procedures use differential centrifugation to separate the intact mitochondria using a bench-top microcentrifuge and are completed in less than 60 minutes. In addition, both procedures have been optimized for maximum yield of mitochondria with minimal damage to integrity. The isolated mitochondria may be used for a number of downstream applications, including 1D and 2D Western blotting and protein profiling using mass spectroscopy.
Highlights:
Quick and Convenient – isolate intact mitochondria in less than 60 minutes
Versatile – offers two methods of isolation from both soft and hard tissue samples
Multi-sample format – reagent-based approach enables simultaneous processing of multiple samples
Optional alternate method – reagents and protocol included for the traditional Dounce homogenization procedure with fewer required strokes
Bench-top compatible – both procedures can be performed using a microcentrifuge tube
Product Details:
COX4
VDAC
Cytochrome C
M
C
M
C
M
C
Reagent-based
Dounce-based
Integrity of mitochondria extracted from tissues. Mitochondria (M) and cytosolic (C) fractions were prepared from fresh rat liver (Panels 1, 2, 3, 5 and 6) and heart (Panel 4) tissue samples using the reagent-based and Dounce homogenization methods of the Thermo Scientific Mitochondria Isolation Kit for Tissue. Fractions were analyzed via Western blot for COX4, VDAC and Cytochrome C mitochondrial proteins. Thermo Scientific SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080) was used for detection. COX4 is an inner-mitochondria membrane protein, VDAC is an outer-mitochondria membrane protein and cytochrome C is located in the intermembrane space. Results indicate that cytosolic proteins were thoroughly separated from the isolated mitochondria.
References:
Zhang, Q., et. al. (2010). Circulating Mitochondrial DAMPs Cause Inflammatory Responses to Injury. Nature. 464(7285): 104-107.
