MDA-MB-231 cells were seeded on top of either compliant (1kPa) or stiff (10kPa) gels for 24 hours and treated with or without the FAK inhibitor PF573228 (MilliporeSigma). Cells were rinsed with 1X PBS and lysed with 4X SDS sample buffer (4X Tris-Cl/SDS, pH6.8, 30% v/v glycerol, 10% w/v SDS, 0.09% v/v 2-mercaptoethanol, and 0.012% w/v Bromophenol Blue). Standard SDS-PAGE was conducted usingBio-Rad Any kD Mini-PROTEAN (4569035; Bio-Rad gels and PVDF membranes (Bio-rad). Membrane washing steps were performed with 0.1% polyoxyethylene 20 sorbitan monolaurate (Tween; JT Baker, Phillipsburg, NJ) in Tris-buffered saline. Blocking was performed with 5% milk in the washing buffer. Primary antibodies (GAPDH Biolegend poly6314; CSF-1 Santa Cruz sc-365779) were diluted in blocking buffer at 1:1000 dilution and applied to the membranes overnight at 4°C. Horseradish-peroxidase conjugated secondary antibodies were applied to the membranes in blocking buffer at 1:2000 dilution for 1 hour at room temperature. Membranes were imaged using SuperSignal chemiluminescent substrate and a FujiFilm ImageQuant LAS-4000. Quantification of protein expression was normalized to GAPDH loading control and densitometry was performed using Fiji.
Sc 365779
Manufactured by Santa Cruz Biotechnology
Sc-365779 is a laboratory product from Santa Cruz Biotechnology. It is a research-grade tool intended for use in scientific experiments and investigations. The core function of this product is to facilitate various laboratory procedures and analyses. No further details on the intended use or specifications of Sc-365779 can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sc 365779
1
Mechanosensitive Signaling in TNBC Cells
2
Mechanosensitive Signaling in TNBC Cells
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MDA-MB-231 cells were seeded on top of either compliant (1kPa) or stiff (10kPa) gels for 24 hours and treated with or without the FAK inhibitor PF573228 (MilliporeSigma). Cells were rinsed with 1X PBS and lysed with 4X SDS sample buffer (4X Tris-Cl/SDS, pH6.8, 30% v/v glycerol, 10% w/v SDS, 0.09% v/v 2-mercaptoethanol, and 0.012% w/v Bromophenol Blue). Standard SDS-PAGE was conducted usingBio-Rad Any kD Mini-PROTEAN (4569035; Bio-Rad gels and PVDF membranes (Bio-rad). Membrane washing steps were performed with 0.1% polyoxyethylene 20 sorbitan monolaurate (Tween; JT Baker, Phillipsburg, NJ) in Tris-buffered saline. Blocking was performed with 5% milk in the washing buffer. Primary antibodies (GAPDH Biolegend poly6314; CSF-1 Santa Cruz sc-365779) were diluted in blocking buffer at 1:1000 dilution and applied to the membranes overnight at 4°C. Horseradish-peroxidase conjugated secondary antibodies were applied to the membranes in blocking buffer at 1:2000 dilution for 1 hour at room temperature. Membranes were imaged using SuperSignal chemiluminescent substrate and a FujiFilm ImageQuant LAS-4000. Quantification of protein expression was normalized to GAPDH loading control and densitometry was performed using Fiji.
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