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Solution Informationhelp
Enzyme: G-protein coupled receptor 55
inhibitor: BDBM75871
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 GPR55 receptor 3) Culture Media: MEM with L-glutamine, Pen-strep, 10% Fetal Bovine Serum and selection antibiotics - 100ug/ml G418 and 50ug/ml Zeocin 4) Agonist Working Solution: Lysophosphatidylinositol (Avanti Polar #850090P, 5mM stock in DMSO) diluted to 50uM in water. 5) Control Working Solution: 100% DMSO (for positive control wells: no agonist will be added to mimic 100% inhibition of agonist-mediated response; for negative control wells: LPI agonist will be added to mimic 0% inhibition of agonist-mediated response). 6) Test Compounds Working Solution: 10mM 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 (10mM TRIS, 10mM EDTA, 100mM NaCl, pH 7.4). Assay Procedure: 1) 45 ul of cell suspens
 
 

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Last update November 1, 2007
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