Solution Informationhelp
Enzyme: Kappa-type opioid receptor
inhibitor: BDBM54818
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 (Greiner# 781091) 2) U2OS (Human Osteosarcoma) cell line stably expressing the Beta-arrestin GFP and the KOR receptor 3) Culture Media: MEM with L-glutamine, Pen-strep, 10% Fetal Bovine Serum and selection antibiotics - 200ug/ml G418 and 100ug/ml Zeocin 4) Agonist Working Solution: Dynorphin (American Peptide #24-4-50, 5mM stock in DMSO) diluted in water to 50uM. 5) DMSO Control Working Solution: 3.1% DMSO in water 6) Fixative Working Solution: 6% Paraformaldehyde (PFA) diluted in PBS. 7) Nuclear Stain Working Solution: DAPI (Invitrogen, D1306) diluted to 150ng/ml in DAPI buffer (10mM TRIS, 10mM EDTA, 100mM NaCl, pH 7.4). Dose Response Assay Procedure: 1) 45ul of cell suspension (200,000 cells/ml in culture medium) was dispensed in each well of the assay plates using a Wellmate bulk dispenser. 2) Plates are incubated overnight or approximately 20 hours at 37 degree C and 5% CO2. 3) Serum is removed by media aspiration a
 
 
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