Solution Informationhelp
Enzyme: Cannabinoid receptor 2
inhibitor: BDBM68296
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Assay Materials: 1) 384-well plates, black with clear bottom (Aurora Biosciences # 032451) 2) U2OS (Human Osteosarcoma) cell line stably expressing the Beta-arrestin GFP and the CB2 receptor 3) Culture Media: MEM with L-glutamine, Pen-strep, 10% Fetal Bovine Serum and selection antibiotics - 200 ug/ml G418 and 100 ug/ml Zeocin 4) Positive Control Working Solution: WIN55212 (Sigma W102 25mg) - 5 mM stock in DMSO) diluted to 2.75 uM in water. 5) Negative Control Working Solution: AM251 (Sigma A6226-10 mg) - diluted to 11 uM in water. 6) Test compounds from dry powders working solution: 10 mM in 100%DMSO 7) Fixative Working Solution: 6% Paraformaldehyde (PFA) diluted in PBS. 8) Nuclear Stain Working Solution: DAPI (Invitrogen, D1306) diluted to 150ng/ml in DAPI buffer (10 mM TRIS, 10 mM EDTA, 100 mM NaCl, pH 7.4). Hit Confirmation Procedure: 1) 45 ul of cell suspension (200,000 cells/ml in culture medium) was dispensed in each well of the assay plates using a Wellmate bulk dispenser.
 
 
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