For plate colony assay, 500 to 1000 cells per well (cell line dependent) were seeded in six-well plates in 2 ml of media and resupplied with fresh media every week. After 2 to 3 weeks, cells were fixed with 10% (w/v) formaldehyde for 15 min and stained with 0.05% (w/v) crystal violet supplemented with 10% ethanol and 10% methanol for 20 min at room temperature. The plates were then washed with distilled water, air dried, and scanned using a Canon scanner. The cell colony numbers were then manually counted. Soft-agar assays were performed as previously described (19 (link)). When applicable, AKT inhibitors MK2206 (#S1078, Selleckchem) and GSK1411795 (#S7492, Selleck), and mTOR kinase inhibitors PP242 (#S2218, Selleckchem) and INK128 (#HY-13328, MedChemExpress) were added during setup and added with fresh media every week.
Gsk1411795
Manufactured by Selleck Chemicals
GSK1411795 is a chemical compound used in laboratory research. It functions as a selective inhibitor of the enzyme PI3K (phosphoinositide 3-kinase).
Automatically generated - may contain errors
2 protocols using gsk1411795
1
Plate Colony and Soft-Agar Assays with Kinase Inhibitors
2
Colorimetric Colony Formation Assay
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For plate colony assay, 500-1000 cells/well (cell line dependent) were seeded in 6-well plates in 2 ml media and replaced with new media every week. After 2-3 weeks, cells were fixed with 10% (w/v) formaldehyde for 15min and stained with 0.05% (w/v) crystal violet supplemented with 10% ethanol and 10% methanol for 20 min at room temperature. The staining solution was then removed and wells washed with distilled water. The plates were then air-dried and scanned using a Canon scanner, and colony numbers were calculated. Soft agar assays were performed as previously described (19) . AKT inhibitors MK2206 (Selleck #S1078) and GSK1411795 (Selleck #S7492) were added during setup and replaced with fresh media every week.
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