PBI-Shredder O2k-Set
Description | PBI-Shredder HRR-Set: Auxiliary HRR-Tool for tissue homogenate preparation; the Shredder-Kit Box contains the heavy duty high torque SG3 driver with convertible handle, SG3 base with 3 position force setting lever (FSL), battery charger, two lithium ion batteries, Shredder-Tube Cap Tool and Shredder-Tube Ram Tool. The PBI-Shredder HRR-Set includes the Shredder-Kit Box with 1 box of 100 Shredder-Tubes FT500-PS, 1 box of 100 Shredder-Tubes FT500-PMS100, a pair of sharp forceps for tissue dissection and a pair of scissors.
OROBOROS INSTRUMENTS: world-wide distributor. |
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Product ID | 13200-04 |
Type | O2k, PBI-Shredder, Catalogue |
Link | PBI-Shredder @OROBOROS, Manual_PBI-Shredder, O2k-Catalogue: PBI-Shredder, Purchase Order @OROBOROS, Shredder vs. Fibres |
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PBI-Shredder HRR-Set consists of
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- >>> Go to O2k-Catalogue OROBOROS, or Order @OROBOROS
Instructions
Using the Shredder-Tube Cap Tool, a serrated [[Shredder-Ram}} is inserted into one of the Shredder-Tubes, pressing the sample between the serratged surface of the Shredder-Ram and the lysis disk of the tube. Ice-cold mitochondrial medium (200-700 Β΅l MiR06 or MiR06Cr) is then added to the opposite Cap side of the tube, which is closed by a Shredder-Screw Cap, again using the Shredder-Tube Cap Tool. The filled Shredder-Tube is placed into the pre-cooled Shredder Base, with the Ram side facing downwards, and the SG3 driver is placed onto the Cap. The force setting lever is placed into the position (1-low, 2-medium), and shredding for 20-30 s is sufficient for several different tissues. The processed homogenate is sampled from the Shredder-Tube by unscrewing the Shredder-Screw Cap using the Shredder-Tube Cap Tool and removing the cap, for transfer of the sample into the O2k-Chamber.
Advantages of the PBI-Shredder
- Low shear mechanical device for gentle, rapid and safe disruption of tissues;
- Three position lever for setting reproducible force to the sample during the shredding process;
- Gentle enough for isolating intact, functional mitochondria;
- Powerful enough to rapidly break apart difficult samples;
- Standardized preparations of high quality;
- Enables reproducible results;
- Easy handling, especially for beginners;
- Processing containers:
- > Standard tubes for ambient pressure processing
- > Closed containers help to ensure safety throughout the entire sample preparation process;
- > Excellent for collection, storage, transport and processing
- High quality of functional mitochondria is obtained from skeletal muscel and kidney as evaluated by high resolution respirometry:
Traditional methods for homogenization
- Homogenizer-based protocols that require extensive operator experience (highly skilled personnel) to obtain reproducible high-quality preparations
- These methods limit dissemination, impede scale-up, and contribute to difficulities in reproducing experimental results over time and across laboratories
- During manual homogenization it is difficult to know when sufficient tissue disruption has occured - overhomogenization!
- The current protocols rely on Dounce, Potter-Elvehjem, and rotor/stator shearing homogenization:
- Pallotti F, Lenaz G (2007) Isolation and subfractionation of mitochondria from animal cells and tissue culture lines. Methods Cell Biol 80: 3-44.
- Frezza C, Cipolat S, Scorrano L (2007) Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nat Protoc 2: 287-295.
- Kaufmann P, TΓΆrΓΆk M, Zahno A, Waldhauser KM, Brecht K, KrΓ€henbΓΌhl S (2006) Toxicity of statins on rat skeletal muscle mitochondria. Cell Mol Life Sci 63: 2415-2425.