From Bioblast
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| 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. The PBI-Shredder HRR-Set includes the Shredder-Kit Box with 100 Shredder-Tubes, 100 Shredder-Tubes\Metal, a pair of sharp forceps for tissue dissection and a pair of scissors.
OROBOROS INSTRUMENTS: world-wide distributor. |
|---|---|
| Product ID | 13200-02 |
| 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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Contents |
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PBI-Shredder HRR-Set consists of
| Title | Description | Product id | Product image |
|---|---|---|---|
| PBI-Shredder SG3 | PBI-Shredder SG3 for tissue homogenate preparation, heavy duty high torque SG3 driver with convertible handle, SG3 base with 3 position force setting lever (FSL), battery charger and two lithium ion batteries. The PBI-Shredder SG3 is included in the PBI-Shredder HRR-Set. Select 230 V or 120 V. OROBOROS INSTRUMENTS: world-wide distributor. | 52100 |  |
| Shredder-Kit Box | Shredder-Kit Box: box for storage and shipping, for PBI-Shredder SG3 | 52101-01 | |
| Shredder-Tube Cap Tool | Shredder-Tube Cap Tool: component of PBI-Shredder_HRR-Set. | 52130-01 |  |
| Shredder-Accessory Box | Shredder-Accessory Box: 71x335x240 mm inner dimensions, for storage and shipping of Shredder accessories. | 52201-01 | |
| Shredder-Tubes | Shredder-Tubes: consisting of Shredder Tube FT500-PS with Lysis Disk, serrated Shredder-Ram and Shredder-Screw Cap, coral colour (Box of 100). 1 box is included in the PBI-Shredder HRR-Set. | 52210-01 |  |
| Shredder-Tubes\Metal | Shredder-Tubes\Metal: consisting of Shredder Tube FT500-PMS with Metal Lysis Disk, serrated Shredder-Ram and Shredder-Screw Cap, coral colour (Box of 100). 1 box is included in the PBI-Shredder HRR-Set. | 52220-01 | |
| Forceps\stainless Steel\straight Tip\sharp | Forceps\stainless Steel\straight Tip\sharp: for tissue preparation, stainless steel, antimagnetic. One pair is recommended for insertion of the sample into the O2k-Chamber and for handling in combination with Forceps\stainless Steel\rounded Tip\sharp. Set: in HRR-Dissection Set and PBI-Shredder HRR-Set. | 54210-01 |  |
| Scissors\stainless steel\straight Tip\sharp | Scissors\stainless steel\straight Tip\sharp, included in PBI-Shredder HRR-Set and HRR-Dissection Set. | 54310-01 |  |
- >>> 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 or Shredder-Tubes\Metal, pressing the sample between the serratged surface of the Shredder-Ram and the lysis disk of the tube. Ice-cold mitochondrial medium (200-700 µl MiR06 or MiR07) 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, Török M, Zahno A, Waldhauser KM, Brecht K, Krähenbühl S (2006) Toxicity of statins on rat skeletal muscle mitochondria. Cell Mol Life Sci 63: 2415-2425.
