The Published Kinase Inhibitor Set (PKIS) was acquired from GlaxoSmithKline by the University of Texas Southwestern High Throughput Screen facility. We modified the PKIS by adding torin1 and rapamycin into our screen. 100μl HCEC (1,000 cells/ml) cells were seeded into each wells and let adhere O/N. Half the plates received fresh media with 1% DMSO, the other half received fresh media with 300 μM BIO. Library compounds and controls were added using Echo liquid handler (Labcyte). 6 doses of each drug at 10,000 3,000, 1000, 300, 100, 30 nM were tested. Cells were then incubated for 72 hrs at 37°C and 5% CO2. Next, media was removed and 25 μl of CellTiter-Glo diluted 1:5 with passive lysis buffer (Promega) was added. Plates were incubated for 10 mins at RT with shaking and read on Envision (Perkin Elmer).
Published kinase inhibitor set
Manufactured by GlaxoSmithKline
The Published Kinase Inhibitor Set is a collection of small-molecule compounds known to inhibit the activity of various kinases. The set provides researchers with a diverse selection of well-characterized kinase inhibitors for use in biochemical and cellular assays.
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2 protocols using published kinase inhibitor set
1
High-Throughput Kinase Inhibitor Screening
2
High-Throughput Kinase Inhibitor Screening
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The Published Kinase Inhibitor Set (PKIS) was acquired from GlaxoSmithKline by the University of Texas Southwestern High Throughput Screen facility. We modified the PKIS by adding torin1 and rapamycin into our screen. 100μl HCEC (1,000 cells/ml) cells were seeded into each wells and let adhere O/N. Half the plates received fresh media with 1% DMSO, the other half received fresh media with 300 μM BIO. Library compounds and controls were added using Echo liquid handler (Labcyte). 6 doses of each drug at 10,000 3,000, 1000, 300, 100, 30 nM were tested. Cells were then incubated for 72 hrs at 37°C and 5% CO2. Next, media was removed and 25 μl of CellTiter-Glo diluted 1:5 with passive lysis buffer (Promega) was added. Plates were incubated for 10 mins at RT with shaking and read on Envision (Perkin Elmer).
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