Transwell migration assays were performed, as previously described (12 (link)). Briefly, the cells were seeded into a 24-well Transwell plates (Millipore, Bedford, MA, USA) in 10% FBS medium at a density of 1×105 cells/well. Following incubation for 24 h at 37°C, the medium was replaced with serum-free medium and the cells were treated with 20, 40 or 80 µM of luteolin or 300 nM Taxol for 24 h at 37°C, while RPMI-1640 medium, containing 10% FBS was added to the lower chamber. The NCI-H460 cells were treated with the drug and cultured at 37°C for a further 5 h. The non-adherent cells were removed by washing with PBS, and the adherent cells were fixed in ethanol. Following staining with 0.1% crystal violet (Tokyo Chemical Industry, Tokyo, Japan), images were captured by microscopy (IX81; Olympus, Tokyo, Japan).
Crystal violet
Manufactured by Tokyo Chemical Industry
Sourced in Japan
Crystal violet is a synthetic dye commonly used as a staining agent in microscopy and biology laboratories. It is a water-soluble, dark purple crystalline solid that can be used to stain various biological samples, including cells and tissues, for visualization and analysis purposes. The core function of crystal violet is to provide a distinctive color contrast that helps in the identification and differentiation of cellular structures and components under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
24 well transwell plate, by Merck Group (1 mentions)
Matrigel, by Corning (1 mentions)
Paraformaldehyde (pfa), by Biosesang (1 mentions)
Eclipse ts2, by Nikon (1 mentions)
Nile red, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Oxacillin, by Merck Group (1 mentions)
Erythromycin, by Merck Group (1 mentions)
Sypro red, by Merck Group (1 mentions)
3 protocols using crystal violet
1
Transwell Migration Assay for Cell Invasion
2
Cell Migration and Invasion Assay
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Costar Transwell® chambers containing 8.0 μm polycarbonate membrane (CLS3422; Corning) were used to analyze cell migration and invasion. HeLa and HaCaT cells were collected and suspended in 100 μl of serum-free DMEM at a density of 5 × 103 cells/ml and 1.5 × 105 cells/ml, respectively, to analyze migration. HeLa cells were also collected and suspended in 100 μl of the same medium, at a density of 5 × 105 cells/ml, to analyze invasion. Medium supplemented with 10% FBS was subsequently added to the wells. Prior to seeding for the invasion assay, a thin layer of Matrigel (354234; Corning) was added to the membrane matrix, and then 100 μl of prepared cells were added on top of the inserts and Matrigel-coated inserts. The chamber plate was incubated at 37°C and 5% CO2 for 24 h. Invaded cells were fixed with 4% paraformaldehyde (Biosesang, Korea) in phosphate-buffered saline for 30 min, and stained with 0.5% crystal violet (Tokyo Chemical Industry, Japan) for 20 min. Cells were observed and imaged using a microscope Eclipse Ts2 (Nikon, Japan).
3
Characterization of Biomolecular Interactions
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Oxacillin, erythromycin, gentamycin, glutaraldehyde, fluorescein isothiocyanate-labeled concanavalin A (FITC-ConA), SYPRO red, Nile red, and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). 9-Fluorenylmethoxycarbonyl (Fmoc) amino acids and Oxyma pure were purchased from CEM Co. (Matthews, NC, USA). Diisopropylcarbodiimide (DIC) and crystal violet were obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). All other chemicals and solvents were of analytical or reagent grade and used as received.
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