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BD Biosciences Discovery Labware
Human PGP-ATPase Assay
Summary | Materials
| Solutions | Assay
Procedure
Materials
Equipment which will be needed:
| Major
Equipment |
Other
Equipment |
| Analytical balance |
Pipettemen (or equivalent) |
| 37 °C incubator |
Multichannel pipetter |
| Microplate Spectrophotometer, capable of
reading optical density between 630 and 850nm |
|
Materials and supplies needed:
In this section we provide a comprehensive list of material and supplies
which will be needed to perform the assays. (We provide grade or suggested
suppliers with catalog numbers).
| Supplies |
| 96-well microtiter plates |
BD Falcon™ Products
|
| Reagent Reservoirs, 50ml |
|
| Pipette tips |
|
| 20, 200, 1000, and 5000uL Pipettors with
corresponding tips |
|
| Multichannel Pipettor (20-200uL) |
|
Chemicals
| |
| Human Pgp (MDR1) Membranes
|
Gentest 453228 (Old K228) |
| Control Membrane Preparation
for ABC Transporters |
Gentest 453200 (Old K200) |
| Potassium Phosphate, Monobasic
(KH2PO4) |
Baker 4008-05 |
| Potassium Phosphate, Dibasic (K2HPO4)
|
Baker 3252-05 |
| Mes (2-[N-Morpholino] ethanesulfonic
acid) Hydrate |
Sigma M-8250 |
| EGTA (Ethylene Gycol-bis (beta-Aminoethylether)-
N,N,N’,N’-Tetraacetic acid) |
Sigma E-4378 |
| DTT (DL- Dithiothreitol) |
Sigma D-5545 |
| Potassium Chloride |
Baker 3040-01 |
| Tris Base |
Baker 4109-06 |
| Sodium Azide |
Sigma S-2002 |
| Zinc Acetate, Dihydrate |
Sigma Z-4540 |
| L-Ascorbic Acid |
Sigma A-1417 |
| Sodium Hydroxide |
Sigma S-0899 |
| Ammonium Molybdate Tetrahydrate |
Sigma A-7302 |
| (+/-)-Verapamil Hydrochloride |
Sigma V-4629 |
| Sodium Orthovanadate |
Sigma S-6508 |
| ATP (Adenosine 5’-Triphosphate, Magnesium
salt) |
Sigma A-9187 |
| Antifoam A Concentrate |
Sigma A-5633 |
Lauryl Sulfate (SDS; Sodium dodecylsulfate)
Purity:approx 99%, Phosphate: <10 ppm |
Sigma L-3771 |
The Vendors/Catalog numbers listed here are those used in
development of this assay. Similar reagents may be available from other
vendors.
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