ABCD ABCD - Molecular Devices
ABCD ABCD - Molecular Devices
ABCD ABCD - Molecular Devices
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GPCR X1: assay principle<br />
<strong>ABCD</strong><br />
agonist<br />
GPCR X1<br />
α<br />
IP3<br />
PLC<br />
Fluo[ ] Fluo[ ]<br />
Fluo[ ]<br />
Fluo[ ]<br />
Fluo[ ]<br />
Ca 2+ Ca 2+<br />
Ca 2+ Ca 2+<br />
Ca2+<br />
Ralf Heilker<br />
Dep. of Integrated Lead Discovery<br />
Protocol for automated assay<br />
<strong>ABCD</strong><br />
• seed 20 ul/well cell suspension (7.000 cells/well) in serum-free<br />
medium in collagen-coated 384-well plate (1 day before)<br />
• add 20 ul/well <strong>Molecular</strong> <strong>Devices</strong> Ca3 Assay Kit<br />
• incubate for 80 min at room temperature (no lid)<br />
• add 20 ul/well Hank's solution<br />
• dilute single-use compound plate, add "high/low" controls to<br />
columns 23/24<br />
• transfer 20 ul/well diluted compound/control solutions from<br />
compound MTP to assay MTP in FLIPR<br />
Ralf Heilker<br />
Dep. of Integrated Lead Discovery
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