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Core Services
This Core facility is established to provide highly-purified plasma membrane, mitochondrial and microsomal specimens for use in transport, metabolic and biochemical studies (over the Center Grant's next 5-year period). In addition, it will provide a high level of expertise and consultation for the design and interpretation of transport experiments, as well as development of new isolation methods.
At its inception, the Core will be equipped and staffed to provide highly-purified sinusoidal and canalicular plasma membrane vesicles from rat livers with well-established methods. Methodology for isolation of vesicles from human livers will be implemented as the need arises. The facility will be capable of preparing hepatic sinusoidal and canalicular membrane vesicles at both high-volume and low-volume scales (starting with a total of ~150 g or ~30 g liver tissue). A high-volume preparation typically yields sufficient vesicles for ~50-100 sinusoidal and ~25-50 canalicular binding/transport studies (i.e. individual incubations corresponding to 35-50 mg protein). It is estimated that a high-volume preparation obtained once every 1-2 weeks would be sufficient to keep up with the on-going and foreseeable demand. Additional preparations will be made, as needed. the low-volume preparation will be run on-demand, at periods when the need may not justify high-volume preparations.
Costs
The Core facility will provide, at no charge to the investigators, the manpower and the expertise to prepare and test for purity and integrity of membrane vesicles before turning them over to the investigators.
The investigators will be expected to cover the costs of procuring the animals and tissues, as well as the nominal costs of the materials used (chemicals, supplies, etc.).
The costs of animals and materials for sinusoidal and canalicular membrane vesicle preparation and testing on a high-volume scale are estimated to be about $500 per preparation.
The investigators may choose to pay on the basis of obtaining whole preps or partial preps when these are made on an on-going basis.
Breakdown of costs for billing purposes
The following are the costs of materials and consumable for sinusoidal and canalicular membrane vesicles, per preparation, on a high-volume scale. These include costs for preparing and testing for integrity of a batch of vesicles that provide ~50-100 sinusoidal and ~25-50 canalicular individual membrane vesicle incubations (corresponding to 35-50 mg protein per incubation) for binding/transport studies (the costs of the latter not reflected below). Cost for preparations on small-scale can be approximated using the costs per incubation, projected from the large-volume preparation.
| Materials Costs |
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| rate (12 at 150-200g) |
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$150.00
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| sucrose (4 kg) |
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$90.00
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| ATP (2.5 g) |
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$50.00
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| miscellaneous chemicals |
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$10.00
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| disposable cubettees (100) |
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$20.00
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| 3H-alanine (~100 Ci) |
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$20.00
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| 3H-tuarocholic acid (~10 Ci) |
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$30.00
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| membrane filters (100) |
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$40.00
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| scintillation cocktail (4 liters) |
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$60.00
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| scintillation vials (100) |
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$30.00
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| Total costs |
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$500.00
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