PDAC organoids were dissociated and 3 × 103 cells were seeded in 150 μl of 90% human organoid feeding media supplemented with 10.5 μM Y27632 (Selleckchem) and 10% growth factor reduced Matrigel (Corning) onto 96-well clear flat bottom plates (Corning) coated with poly(2-hydroxyethyl methacrylate) (Sigma-Aldrich). On the following day, organoids were drugged with CHK1i (prexasertib, 2 to 32 nM), ERKi (SCH772984, 0.008 to 2 μM) and chloroquine (3.25 μM) using a Tecan D300e digital dispenser. Ten days after drugging, organoids were imaged with a Molecular Devices SpectraMax i3x MiniMax 300 imaging cytometer. After image acquisition, organoid viability was assessed with the CellTiter-Glo 3D Cell Viability Assay (Promega) on a SpectraMax i3x plate reader, according to the manufacturer’s protocol.
Spectramax i3x plate reader
Manufactured by Promega
The SpectraMax i3x is a multi-mode microplate reader capable of absorbance, fluorescence, and luminescence detection. It features a monochromator-based optics system and a high-sensitivity PMT detector. The SpectraMax i3x can be used for a variety of applications in life science research and drug discovery.
Automatically generated - may contain errors
Lab products found in correlation
Clear flat bottom 96 well plate, by Corning (2 mentions)
Spectramax i3x minimax 300 imaging cytometer, by Molecular Devices (2 mentions)
Celltiter glo 3d cell viability assay, by Promega (2 mentions)
Poly 2 hydroxyethyl methacrylate, by Merck Group (2 mentions)
Y 27632, by Selleck Chemicals (2 mentions)
Matrigel, by Corning (2 mentions)
Camp glo assay, by Promega (1 mentions)
3 protocols using spectramax i3x plate reader
1
PDAC Organoid Viability Assay
2
Intracellular cAMP Modulation in Glia
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Astrocytes and microglia were exposed to 0.75, 1.5, 6, 12 and 24 µM DON for 1 h, and intracellular cAMP concentrations were measured using the cAMP-Glo assay according to the manufacturer‘s guidelines (Promega) using the Spectramax i3x plate reader. The cAMP agonist forskolin (4 µM; 15 min exposure) was used as positive control.
3
PDAC Organoid Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PDAC organoids were dissociated and 3 × 103 cells were seeded in 150 μl of 90% human organoid feeding media supplemented with 10.5 μM Y27632 (Selleckchem) and 10% growth factor reduced Matrigel (Corning) onto 96-well clear flat bottom plates (Corning) coated with poly(2-hydroxyethyl methacrylate) (Sigma-Aldrich). On the following day, organoids were drugged with CHK1i (prexasertib, 2 to 32 nM), ERKi (SCH772984, 0.008 to 2 μM) and chloroquine (3.25 μM) using a Tecan D300e digital dispenser. Ten days after drugging, organoids were imaged with a Molecular Devices SpectraMax i3x MiniMax 300 imaging cytometer. After image acquisition, organoid viability was assessed with the CellTiter-Glo 3D Cell Viability Assay (Promega) on a SpectraMax i3x plate reader, according to the manufacturer’s protocol.
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