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 consultaton 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. Mehtodology 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.
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 weel 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 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 memberane 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.