Minute™ Mitochondrial Membrane Protein Isolation Kit

Invent Biotechnologies Minute™ Mitochondrial Membrane Protein Isolation Kit is composed of optimized buffers and protein extraction filter cartridges with 2.0 ml collection tubes. The kit is designed to rapidly isolate native mitochondrial proteins from cultured mammalian cells or tissues. Due to the use of protein extraction filter cartridges, the protein isolation procedure is simple, easy and user friendly with high yield. Unlike many commercial mitochondrial preparation kits, this kit offers wide range of starting cells (5-40 millions/sample) and intact mitochondria are isolated. The buffers are detergent and EDTA free. A Dounce homogenizer or a tissue blender is not needed. The procedure can be completed in about 30 min.

Applications
The kit is designed to rapidly isolate mitochondria membrane proteins from cultured cells or tissues for applications such as SDS-PAGE, immunoblottings, ELISA, IP, membrane protein structure analysis, 2-D gels, enzyme activity assays and other applications.
Buffer Formulations: Proprietary

Enrichment of Mitochondrial Marker (Ubiquinol-Cytochrome C Reductase Core Protein) with Minute™ Mitochondria Isolation Kit.
A. SDS-PAGE profiles of total protein extract vs. isolated mitochondrial proteins. Lanes 1, 3, 5: total proteins from isolated mouse kidney cells, mouse skeletal muscle and liver tissues respectively. Lanes 2, 4, 6: isolated mitochondrial proteins from mouse kidney cells, mouse skeletal muscle and liver tissues respectively.
B. Western blotting. Proteins in A were transferred to nitrocellulose membranes and probed with anti-ubiquinol-cytochrome C reductase core protein and anti-Lamin B1.
C. Densitometry measurement of mitochondrial marker signals.

Kit components
1. 25 ml buffer A
2. 30 ml buffer B
3. 50 protein extraction filter cartridges
4. 50 collection tubes with cap
5 Plastic rods (4)
6. Tissue dissociation beads


Important Information:
1. Read the entire procedures carefully. Thaw buffer A and buffer B completely, invert the bottles a few times and place on ice. Chill protein extraction filter cartridge with collection tube on ice prior to use.
2. All centrifugation steps should be performed at 4oC in a cold room or in a refrigerated mirocentrifuge.
3. To study protein phosphorylation, phosphatase inhibitors should be added to buffer A prior to use. The use of protease inhibitor cocktails is optional.
4. It is recommended to use BCA Protein Assay Kit for determination of protein concentration.

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