SLC_Transporter_cell_based_assay【DOC精选】.docVIP

Title : SLC Transporter Cell-Based Assay Signature Date Drafter Year / Month / Day Manager Year / Month / Day Issued on : Year / Month / Day Voided on : Year / Month / Day The Original Copy Stored at : Distributed on : Year / Month / Day List of Distribution : No. Distributed to : 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 SOP Revision Record Revision Date Version Revision Details Year/Month/Day 01 Year/Month/Day Year/Month/Day Year/Month/Day SLC Transporter Cell-Based Assay Described as follows is the summary of method in SLC Transporter Cell-Based Assay. Stable Cell Lines and Substrates Transporters Cell Lines Plate/# of Cells Radioactive Labeled Substrate Buffer pH Substrate Concentration (Km) Human OCT1 HEK293 12-well Plate/ 5x105cells/ well [Biguanidine-14C]- Metformin Hydrochioride 5 ｍM pH7.4 Human OCT2 [Biguanidine-14C]- Metformin Hydrochioride 2.8 mM Human OATP1B1 [6,7-3H(N)]-Estrone Sulfate, Ammonium salt 78.8 nM Human OATP1B3 [Estradiol-6,7-3H(N)]- Estradiol-7β-D-glucuronide 27.4 μM Human OAT1 p-[GLYCYL-2-3H]- Aminohippuric acid 20.3 μM Human OAT3 [6,7-3H(N)]-Estrone Sulfate, Ammonium salt 19.5 μM Reagents, Materials and Equipments Reagents Reagent Manufacturer Model Number Storage Temperature Light Shielding Expiration Date NaCl Nacalai Tesque 31320-05 room temperature not required 5 yrs after opening KCl Nacalai Tesque 28514-75 room temperature not required 5 yrs after opening D-(+)-glucose Nacalai Tesque 16805-35 room temperature not required 5 yrs after opening CaCl2?2H2O Nacalai Tesque 06731-05 room temperature not required 5 yrs after opening KH2PO4 Nacalai Tesque 28736-75 room temperature not required 5 yrs after opening MgSO4?7H2O Nacalai Tesque 2 room temperature not required 5 yrs after opening HEPES Nacalai Tesque 02443-34 room temperature not required 5 yrs after opening 1N NaOH Nacalai Tesque 37421-05 room temperature r