Solution Information | help | |
Enzyme: | Sphingosine 1-phosphate receptor 1 | |
inhibitor: | BDBM37176 | |
substrate: | n/a | |
Solution Type: | Aqueous | |
pH at Preparation: | n/a | |
Temp. Prep.: | n/a | |
Comments: | For these dose response assays, one of two cell lines was used. The S1P-1 "potentiator" and agonist assays used a Chinese Hamster Ovary (CHO) cell line stably transfected with human S1P1 receptor (pcDNA3.1-S1P1) and a cyclic AMP response element (CRE) reporter construct. The "parental" counter screen dose response assay used a CHO Cre Bla cell line (Invitrogen part#K1129). All cells were cultured in T-175 cm2 Corning flasks (part#431080) at 37 degrees C and 95% RH. The growth media consisted of Dulbecco#s Modified Eagle#s Media (Invitrogen part#11965-092) containing 10% v/v heat inactivated bovine growth serum (Hyclone part#SH30541.03), 0.1 mM NEAA (Invitrogen part#1114-050), 1 mM Sodium Pyruvate (Invitrogen part#11360-070), 25 mM HEPES (Invitrogen part#15630-080), 5 mM L-Glutamine (Invitrogen part#25030-081), and 1x penicillin-streptomycin (Invitrogen part#15140-122).For the parental cell line no selection antibiotic was used but for the S1P1 transformed cells 2 mg/ml Geneticin (Inv | |
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