Cells were diluted to 5.0 x 104 cells per mL, and 20 uL of cell suspension was added to wells of an opaque 384 well plate. After overnight recovery, cells were subjected to a dose response curve of increasing drug concentration in half log steps. For single drug curves, osimertinib (Cayman AZD9291) range was .00003 to 30 uM. For dual drug curves, osimertinib range was .03 to 10 uM, and either KRAS inhibitor (ARS853, Cayman) or FGFR1 inhibitor (PD166866, Cayman) range was .00003 to 30 uM. 10 mM stocks of drugs were made by diluting with DMSO, and half-log drug series were diluted fresh with complete media. Cell viability was read after 48 hours using Promega’s Cell Titer Glo. Luminescence values were normalized to non-drug treated controls, and plotted using Graphpad.
Ars 853
Manufactured by Cayman Chemical
ARS-853 is a chemical compound used for laboratory research purposes. It serves as a core functional component in various experimental setups and analytical procedures. The detailed specifications and technical details of this product are available upon request.
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Lab products found in correlation
3 protocols using ars 853
1
Combinatorial Drug Screening in Cancer
2
Cell Viability Assay for 2D and 3D
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For the drug titration experiments, ~16,000 cells were seeded
into tissue-culture treated 96-well plates in RPMI 1640 growth media (2D
monolayers) or ultra-low attachment 96-well plates in RPMI 1640 growth median
with 0.75% methylcellulose. Cells were then grown for the next 72 hours in
presence of titrated inhibitors. At the 72 hr point, 1/10th volume of alamarBlue
reagent (ThermoFisher, DAL1100) was added to cells and incubated ~2 hours
for 2D monolayer cells and ~10 hours for 3D spheroids at 37°C.
Fluorescence signals were then measured in a fluorescence plate reader (TECAN,
#30016056, excitation at 560 nm, emission at 590 nm) to estimate relative number
of live cells at different dosages of the inhibitors. Wild type H23 cells were
used in the experiments where efficacies of small molecule inhibitors were
compared between 2D and 3D. To test whether CPD deletion sensitizes cells
against ARS-853, H23 cells with Safe-sgRNA and with CPD-sgRNA (no fluorescent
marker) were used. Small inhibitors were obtained from the following sources :
Savolitinib from Selleckchem (S7674), Linsitinib from VWR (# 10189–468),
FURIN inhibitor I from Sigma Aldrich (# 344930), and ARS-853 from Cayman
chemical (# 1629268–00-3).
into tissue-culture treated 96-well plates in RPMI 1640 growth media (2D
monolayers) or ultra-low attachment 96-well plates in RPMI 1640 growth median
with 0.75% methylcellulose. Cells were then grown for the next 72 hours in
presence of titrated inhibitors. At the 72 hr point, 1/10th volume of alamarBlue
reagent (ThermoFisher, DAL1100) was added to cells and incubated ~2 hours
for 2D monolayer cells and ~10 hours for 3D spheroids at 37°C.
Fluorescence signals were then measured in a fluorescence plate reader (TECAN,
#30016056, excitation at 560 nm, emission at 590 nm) to estimate relative number
of live cells at different dosages of the inhibitors. Wild type H23 cells were
used in the experiments where efficacies of small molecule inhibitors were
compared between 2D and 3D. To test whether CPD deletion sensitizes cells
against ARS-853, H23 cells with Safe-sgRNA and with CPD-sgRNA (no fluorescent
marker) were used. Small inhibitors were obtained from the following sources :
Savolitinib from Selleckchem (S7674), Linsitinib from VWR (# 10189–468),
FURIN inhibitor I from Sigma Aldrich (# 344930), and ARS-853 from Cayman
chemical (# 1629268–00-3).
3
Cell Viability Assay for 2D and 3D
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