For cell migration assays, 1 × 105 control or long-term PM2.5 treated A549 cells were transferred into a Transwell insert, and incubated with complete medium for 24 h. Cell migration was detected from triplicates for each treatment after crystal violet staining. For cell invasion assays, 1 × 105 control or long-term PM2.5 treated A549 cells were transferred into the Transwell insert containing wells pre-filled with Matrigel (CytoSelect 24-Well Cell Invasion Assay Kit; Cell Biolabs, USA) and cultured with complete medium for 48 h. Cell invasion was determined from triplicates for each treatment after crystal violet staining.
Cytoselect 24 well cell invasion assay kit
Manufactured by Cell Biolabs
Sourced in United States
The CytoSelect 24-well cell invasion assay kit is a quantitative tool for measuring the invasive potential of cells. The kit provides a defined basement membrane matrix for cells to invade through, allowing for the assessment of their migratory and invasive capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Cytoselect 48 well cell adhesion assay, by Cell Biolabs (2 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
Versamax microplate reader, by Molecular Devices (1 mentions)
24 well transwell, by Corning (1 mentions)
Cyquant gr dye, by Thermo Fisher Scientific (1 mentions)
Infinite m200, by Tecan (1 mentions)
Cell invasion assay kit, by Merck Group (1 mentions)
Transwell insert, by Corning (1 mentions)
Cytation 5, by Agilent Technologies (1 mentions)
Bz x700, by Keyence (1 mentions)
Microplate reader, by Molecular Devices (1 mentions)
G lisa rhoa activation assay biochem kit, by Cytoskeleton (1 mentions)
Nad nadh quantification kit, by Abcam (1 mentions)
Infinite m200 microtiter plate reader, by Tecan (1 mentions)
Versamax, by Molecular Devices (1 mentions)
Imark microplate absorbance reader, by Bio-Rad (1 mentions)
Cell titer cell viability assay, by Promega (1 mentions)
Cytoselect 24 well wound healing assay kit, by Cell Biolabs (1 mentions)
Facscalibur, by BD (1 mentions)
44 protocols using cytoselect 24 well cell invasion assay kit
1
Cell Migration and Invasion Assays
2
Cell Invasion Assay Protocol
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Cell invasion was evaluated with the CytoSelect 24-Well Cell Invasion Assay kit (Cell Biolabs, San Diego, CA). After siRNA knockdown, the cells were cultured in 10−9 M E2-containing medium for 36 h and starved in serum free medium for overnight. Then, 3 × 105 cells in serum-free medium were seeded on a basement membrane-coated insert with 8 μM pores, and 500 μl of medium containing 10% of charcoal-stripped FBS and E2 (10−9 M) were added to the lower chamber. After a 24-h incubation, cells that had penetrated the insert pore were extracted and absorbance at OD560nm was measured.
3
Evaluating Cell Migration and Invasion with IWR-1 Treatment
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HCT116 and HT29 cell migration was analyzed using the in vitro wound healing assay. Cells were grown to confluence in 48-well plates and changed to serum-free medium for additional 24 h. Cell monolayers were scraped with a micropipette tip and treated with 10 uM IWR-1. The wound area was photographed under phase-contrast microscopy before and 24 h after the treatment and the percentage of wound closure was determined as: [(initial area - final area)/initial area] × 100 [64 (link)].
Invasion assays were conducted using the CytoSelect 24-Well Cell Invasion Assay Kit (Cell Biolabs, San Diego, CA, USA). Briefly, 300 μL of HCT116 and HT29 cells (1 × 105cells/ml) in serum-free medium was plated into the CytoSelect basement membrane chamber, respectively, and 500 μL of 10% FBS-containing RPMI was added to the lower well of the invasion plate; both upper and lower chambers contained the 10 uM IWR-1. The chambers were then incubated for 48 h at 37°C in a 5% CO2 atmosphere, non-migratory cells were removed, and migrated cells were stained, dissociated from the membrane, and their absorbance was measured at 560 nm using the microplate reader (model 680; Bio-Rad, Hercules, CA, USA) [65 (link)].
Invasion assays were conducted using the CytoSelect 24-Well Cell Invasion Assay Kit (Cell Biolabs, San Diego, CA, USA). Briefly, 300 μL of HCT116 and HT29 cells (1 × 105cells/ml) in serum-free medium was plated into the CytoSelect basement membrane chamber, respectively, and 500 μL of 10% FBS-containing RPMI was added to the lower well of the invasion plate; both upper and lower chambers contained the 10 uM IWR-1. The chambers were then incubated for 48 h at 37°C in a 5% CO2 atmosphere, non-migratory cells were removed, and migrated cells were stained, dissociated from the membrane, and their absorbance was measured at 560 nm using the microplate reader (model 680; Bio-Rad, Hercules, CA, USA) [65 (link)].
4
Cell Invasion Assay Protocol
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Cell invasion assays were performed as described previously [31 (link)]. In brief, cells transfected with miRNA mimic or siRNA were seeded at 5 × 105 cells/well in the upper chamber of a CytoSelect 24-Well Cell Invasion Assay kit (Cell Biolabs, San Diego, CA, USA). The number of invading cells was counted in three randomly selected views under a microscope.
5
Cell Invasion Assay Protocol
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Cell invasion was assessed using a CytoSelect 24-well Cell Invasion Assay kit (Cell Biolabs, San Diego, CA, USA) according to the manufacturer’s protocol. Briefly, at 36 hours post-transfection, 1 × 105 cells in 300 μL serum-free medium were added to the upper chamber precoated with basement membrane matrix solution. Subsequently, 0.5 mL of 10% FBS-containing medium were added to the lower chamber as a chemoattractant. The cells were incubated for 36 hours at 37 °C, then the non-invading cells were removed with cotton swabs. The cells, which migrated to the bottom of the membrane, were fixed and stained with staining solution for 10 minutes. The inserts with stained cells were air-dried, then incubated in the extraction solution for 10 minutes and absorbance was measured at a TECAN spectrophotometer (Tecan, Zürich, Switzerland) at 560 nm.
6
Cell Invasion and Migration Assays
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For the cell invasion assays, after 48 h of transfection, 5 × 104 CRC cells were plated at the top of the Matrigel-coated invasion chambers (CytoSelect 24-Well Cell Invasion Assay Kit; Cell Biolabs, U.S.A.) with a serum-free DMEM in six-well plates; 10% FBS DMEM was added to the lower chamber and cultured for 24 h. For cell migration assays, after 48 h of transfection, 5 × 104 cells were plated into the transwell chamber, and cultured with complete medium for 24 h. Finally, all cells were fixed with ethanol and stained with Crystal Violet.
7
Quantifying Cell Invasion Potential
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Cell Invasion was assessed using a CytoSelect 24-well Cell Invasion Assay Kit (Cell Biolabs, Inc., San Diego, CA, USA) with an 8 µm pore size polycarbonate membrane, according to the manufacturer’s instructions. Briefly, after starvation for 24 h in serum-free media, 3.0 × 105 cells that were re-suspended in serum-free media were seeded in the upper chamber. The lower chamber was filled with 500 µL RPMI 1640 medium containing 10% FBS. The plate was incubated at 37 °C in 5% CO2 for 24 h to induce the invasion towards the lower chamber. After removing non-invasive cells from the upper side of the chamber, invaded cells on the reverse side were stained using the provided staining solution. The stained membranes were incubated for 10 minutes with extraction buffer and the lysate was quantified by measuring the absorbance at 560 nm using an IMARK microplate reader (Bio-Rad Laboratories, Inc.).
8
Cell Invasion Assay for Metastatic Potential
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The invasive ability of all tested cell lines was estimated using the CytoSelect 24-well Cell Invasion Assay Kit (Cell BioLabs, Inc., San Diego, CA, United States) according to the manufacturer’s protocol. 1.0 × 105 cells/well were spread in 6-well plates. miRNAs were transfected on the next day. After 24 h of transfection, 3.0 × 105 cells were displaced in the transwell system and incubated for 48 h. The number of invading cells was measured under a microscope after staining with the solution in the kit.
9
Scratch-Induced Differentiation Assay for ESCs
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4 × 105 WT and Smad3−/− ESCs were seeded in six-well tissue culture plates, respectively, in 2i and LIF ESC culture media. After 2 days, the ESC media were changed to differentiation media, in which 2i and LIF were removed and 1 μM RA (Sigma) was added, for another day to promote quick ESC differentiation. When ESCs were completely differentiated into monolayer cells, autoclaved yellow pipette tips were used to generate scratches. After scratching, the detached cells were removed by washing twice with DPBS buffer, and then cultured in differentiation media for another 24 h under normal condition. Images were taken by a Zeiss microscope (Carl Zeiss, Jena, Germany) and analyzed, using Image J software (NIH, Washington, DC, USA), at 0 and 24 h. Triplicate independent assays were performed. The cell invasive assay was performed using the CytoSelect 24-Well Cell Invasion Assay Kit (Cat # CBA-110-COL, Cell Biolabs, Inc., San Diego, CA, USA) according to the manufacturer's instructions.
10
Quantifying Apoptosis and Cell Migration
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For apoptotic cell analysis, PBS- or mJX-594-treated cells were washed with PBS, fixed in 4% paraformaldehyde for 15 min at room temperature, and permeabilized with 0.3% Triton X-100 for 5 min. Apoptotic cells with DNA breaks were detected using the In Situ Cell Death Detection Kit, TMR red (#12 156 792 910; Roche, Indianapolis, IN, USA) according to the supplier’s protocol. For migration assay, cells were treated with PBS or mJX-594 (1 MOI) and plated in the top chamber (5 × 104 cells/well) of 24-well Transwell (8.0 μm pore size; Corning Inc., Corning, NY, USA). Then, a 10% fetal bovine serum-containing medium was placed in the bottom Transwell chamber, and the assembly was incubated at 37 °C for 24 h. In vitro Matrigel invasion assays were performed using CytoSelect™ 24-Well Cell Invasion Assay kit (Cell Biolabs, San Diego, CA, USA) following the manufacturer’s manual. Migrating or invading cells on the bottom surface of the membrane were stained with crystal violet and quantified at OD 560 nm after extraction using the VersaMax microplate reader (Molecular Devices, Sunnyvale, CA, USA).
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