The H358 lung cancer cell line expressing the KRAS G12C alteration was seeded on a collagen-fibronectin matrix in Stacks and treated with 100 nM of adagrasib, a KRAS G12C small molecule inhibitor (Selleck Chemicals, Houston, TX, USA) or with DMSO for 48 hours in serum-free RPMI media. Cells were stained under live conditions with EpCAM Alexa Fluor 647 at 1:50 dilution, Image-iT® Dead™ Green at 1:10,000 dilution, and Hoechst33342 for 60 minutes. Wells were rinsed with PBS, fixed with 1% PFA, washed with PBS, and mounted as described above. Cells were imaged directly in-chip using a Nikon Yokogawa CSU-W1 Spinning Disk Confocal Microscope equipped with Hamamatsu Orca Flash cameras, and the percentage of Image-iT® Dead™-positive cells were quantified using NIS-Elements software. Spot detection function was used to identify individual cells and generate statistics on signal intensity values in each cell and for the sample as a whole. This experiment was performed twice with three technical replicates per experiment.
Kras g12c small molecule inhibitor
Manufactured by Selleck Chemicals
Sourced in United States
The KRAS G12C small molecule inhibitor is a laboratory reagent designed to target the KRAS G12C mutation, a common oncogenic driver in various cancer types. It functions by inhibiting the activity of the KRAS G12C protein, which plays a crucial role in cellular signaling and proliferation. This product is intended for research use only and should not be used for any other purpose.
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2 protocols using kras g12c small molecule inhibitor
1
KRAS G12C Inhibitor Treatment Effects
2
KRAS G12C Inhibition in Lung Cancer
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The H358 lung cancer cell line expressing the KRAS G12C alteration was seeded on a collagen‐fibronectin matrix in Stacks and treated with 100 nM of adagrasib, a KRAS G12C small molecule inhibitor (Selleck Chemicals, Houston, TX, USA) or with DMSO for 48 h in serum‐free RPMI media. Cells were stained under live conditions with EpCAM Alexa Fluor 647 at 1:50 dilution, Image‐iT® Dead™ Green at 1:10 000 dilution, and Hoechst33342 for 60 min. Wells were rinsed with PBS, fixed with 1% PFA, washed with PBS, and mounted as described above. Cells were imaged directly in‐chip using a Nikon Yokogawa CSU‐W1 Spinning Disk Confocal Microscope equipped with Hamamatsu Orca Flash cameras, and the percentage of Image‐iT® Dead™‐positive cells were quantified using NIS‐Elements software. Spot detection function was used to identify individual cells and generate statistics on signal intensity values in each cell and for the sample as a whole. This experiment was performed twice with three technical replicates per experiment.
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