HMC1, HMC3A, and HMC3B cells were seeded in 24-well plates at 800, 600, and 1400 cells/well, respectively. The next day, media was replaced with 500 μL fresh media containing either vehicle (DMSO) or the indicated concentration of drug (BMS-754807 (Sigma-Aldrich #BM0003), PPP (Santa Cruz #SC-204008), SR10221, SR2595 (Sigma-Aldrich #SML2037), or T0070907 (Fisher #NC1015539); final concentration 1% DMSO). Seven days later, media was removed from all wells and cells were fixed in 10% buffered formalin (Fisher #SF994) for 5 min at room temperature. Wells were washed once in ddH2O and then stained in 0.05% crystal violet (Sigma-Aldrich #C6158) for 30 min at room temperature. Wells were then washed an additional 3 times in ddH2O to remove any unbound stain and allowed to dry at room temperature overnight. Once dry, individual wells were imaged at 4X magnification on a BioTek Cytation 5 plate reader (BioTek Instruments, Inc.) and images were stitched using the Gen5 software (v2.09; BioTek Instruments, Inc.) using the default parameters. Colony numbers were quantified manually in ImageJ (Schneider et al., 2012 (link)), where a colony is defined as a cluster of at least 50 individual cells.
Nc1015539
Manufactured by Thermo Fisher Scientific
The NC1015539 is a laboratory equipment product from Thermo Fisher Scientific. It is designed for general laboratory use. The core function of this product is to provide a specific capability or utility within a laboratory setting. No further details about its intended use or capabilities can be provided while maintaining an unbiased and factual approach.
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Lab products found in correlation
2 protocols using nc1015539
1
Quantifying Colony Formation Assay
2
Quantifying Colony Formation Assay
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HMC1, HMC3A, and HMC3B cells were seeded in 24-well plates at 800, 600, and 1400 cells/well, respectively. The next day, media was replaced with 500 μL fresh media containing either vehicle (DMSO) or the indicated concentration of drug (BMS-754807 (Sigma-Aldrich #BM0003), PPP (Santa Cruz #SC-204008), SR10221, SR2595 (Sigma-Aldrich #SML2037), or T0070907 (Fisher #NC1015539); final concentration 1% DMSO). Seven days later, media was removed from all wells and cells were fixed in 10% buffered formalin (Fisher #SF994) for 5 min at room temperature. Wells were washed once in ddH2O and then stained in 0.05% crystal violet (Sigma-Aldrich #C6158) for 30 min at room temperature. Wells were then washed an additional 3 times in ddH2O to remove any unbound stain and allowed to dry at room temperature overnight. Once dry, individual wells were imaged at 4X magnification on a BioTek Cytation 5 plate reader (BioTek Instruments, Inc.) and images were stitched using the Gen5 software (v2.09; BioTek Instruments, Inc.) using the default parameters. Colony numbers were quantified manually in ImageJ (Schneider et al., 2012 (link)), where a colony is defined as a cluster of at least 50 individual cells.
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