In this video, we describe a method highlighting the utility of bioluminescence assay for sensitive and rapid screening of drugs. These drugs can be tested alone or in combination to assess the synergism in their mode of action.
Protocol
1. Bio-luminescence Based Measurement of Cell Viability Coating plates with the extracellular matrix (ECM) mixture (e.g., Matrigel): Add 40 µL of 0.15 mg/mL ECM mixture to each well and incubate the plate for 1 h at 37 °C. Remove the excess ECM mixture and gently rinse once with PBS. Add 100 µL culture medium containing 15,000, 10,000, 8,000, 6,000, 4,000, 2,000, 1,000, and 500 XG387-Luc cells together with 100 µL blank medium as control into each well for 6 repl…
Divulgations
The authors have nothing to disclose.
Materials
B-27
Gibco
17504-044
50X
EGF
Gibco
PHG0313
20 ng/ml
FGF
Gibco
PHG0263
20 ng/ml
Gluta Max
Gibco
35050061
100X
Neurobasal
Gibco
21103049
1X
Penicillin-Streptomycin
HyClone
SV30010
P: 10,000 units/ml S: 10,000 ug/ml
Sodium Pyruvate
Gibco
2088876
100 mM
ABT-737
MCE
Selective and BH3 mimetic Bcl-2 Bcl-xL and Bclw inhibitor
Adavosertib (MK-1775)
MCE
Wee1 inhibitor
Axitinib
MCE
Multi-targeted tyrosine kinase inhibitor
AZD5991
MCE
Mcl-1 inhibitor
A 83-01
MCE
Potent inhibitor of TGF-β type I receptor ALK5 kinase
CGP57380
Selleck
Potent MNK1 inhibitor
Dactolisib (BEZ235)
Selleck
Dual ATPcompetitive PI3K and mTOR inhibitor
Dasatinib
MCE
Dual Bcr-Abl and Src family tyrosine kinase inhibitor
Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening. J. Vis. Exp. (Pending Publication), e20614, doi: (2023).