For cell invasion assay, 1.5 × 105 cells in serum-free 1640 medium were seeded into a matrigel coated chamber (8 μm pore size; Corning Incorporated, NY, USA) and the lower chamber was immediately filled with 500 μL of 1640 medium with 10 % FBS as a chemoattractant. After 24 h of incubation, the non-invading cells were removed from the upper chamber by a cotton swab, and the membranes fixed with methanol and stained by 0.1 % crystal violet. The data are represented as mean ± standard deviation (s.d.), n = 3.
Matrigel coated chamber
Manufactured by Corning
Sourced in United States
Matrigel-coated chambers are a type of laboratory equipment designed to provide a specialized cell culture environment. Matrigel is a gelatinous protein mixture that mimics the extracellular matrix, promoting cell attachment, growth, and differentiation. These chambers offer a controlled and consistent substrate for cell-based experiments and research.
Automatically generated - may contain errors
Lab products found in correlation
Transwell chamber, by Corning (2 mentions)
Microplate reader, by Molecular Devices (2 mentions)
Matrigel, by Corning (2 mentions)
24 well plate chamber insert, by Corning (2 mentions)
Crystal violet, by Merck Group (2 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
96 well plate, by Corning (1 mentions)
Top chamber, by Corning (1 mentions)
24 transwell chamber, by Corning (1 mentions)
Bx41 microscope, by Olympus (1 mentions)
Methanol, by Sangon (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Crystal violet, by Sangon (1 mentions)
Transwell pore polycarbonate membrane insert, by Corning (1 mentions)
Cell counting kit 8 cck 8, by Yeasen (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Inverted microscope, by Leica (1 mentions)
32 protocols using matrigel coated chamber
1
Matrigel-based Cell Invasion Assay
2
Cell Invasion and Motility Assays
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For invasion assays, 1.0 × 105 cells were seeded in a Matrigel-coated chamber with 8.0 mm pores(Corning Inc., NY, USA). For motility assays, 5.0 ×104 cells were plated on uncoated membranes with 8.0 mm pores (Corning Inc.). Cells were seeded in serum-free medium and translocated toward complete growth medium for 24 h.
3
Cell Proliferation and Apoptosis Assay
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Cells transfected with various plasmids were seeded onto 96-well plates (Corning Inc., Corning, NY, USA) at a density of 1 × 104 cells/well in 96-well plates. At different time points (24, 48, and 72 h) after plating, the number of cells was assessed using the Cell Counting Kit 8 according to the protocols of the manufacturer (Dojindo, Tokyo, Japan). The transfected cell lines undergoing apoptosis were distinguished from live and necrotic cells by using Annexin-V and propidium iodide (PI) staining kit (Calbiochem, San Diego, CA, USA). All experiments were independently repeated three times.
For cell invasion assay, 1.5 × 105 cells in serum-free medium were seeded into a Matrigel-coated chamber (8 μm pore size, Corning Incorporated, NY, USA), and the lower chamber was immediately filled with 500 μl of 1640 medium with 10% FBS as a chemoattractant. After 24 h of incubation, the non-invading cells were removed from the upper chamber by a cotton swab. The membranes were fixed with methanol and stained by 0.1% crystal violet. The data were represented as mean ± standard deviation (SD), n = 3.
For cell invasion assay, 1.5 × 105 cells in serum-free medium were seeded into a Matrigel-coated chamber (8 μm pore size, Corning Incorporated, NY, USA), and the lower chamber was immediately filled with 500 μl of 1640 medium with 10% FBS as a chemoattractant. After 24 h of incubation, the non-invading cells were removed from the upper chamber by a cotton swab. The membranes were fixed with methanol and stained by 0.1% crystal violet. The data were represented as mean ± standard deviation (SD), n = 3.
4
Invasion Assay for HCC Cells
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A total of 5 × 104 HCC cells transfected with plasmids were seeded in the upper chamber of a Matrigel-coated chamber (8 μm pore size; Corning) and cultured in serum-free DMEM. After 24 h, the cells with good invasive ability were traversed to the lower surface of the membrane. Then, 4% paraformaldehyde was used to fix the invasive cells, and 0.1% crystal violet was used to stain the invasive cells. Finally, the invasive cells were counted under an inverted microscope.
5
Comprehensive Cell Assays for ccRCC
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Cell proliferation was measured by CCK-8 Kit according to the manufacturer's protocols. For colony formation assay, cells were seeded in 6-well plates at a density of 5 × 102 cells per well then stained and photographed after 2 weeks. Wound healing assay were used to detect cell migration ability as previously describe [24] (link). Cell invasion assay were performed using matrigel-coated chamber (Corning, USA). In brief, 5 × 104 ccRCC cells with serum-free media were seeded into the upper chambers and stained after 24 h.
6
Transwell Invasion Assay with Matrigel
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Transwell assay was conducted using Matrigel coated chamber (Corning, USA). Cells were transfected for 48 hours and harvested to seed into the upper chamber with serum-free medium. Medium containing 10% FBS was added into the lower chamber. After 24 hours, cells invaded through the membrane were fixed with methanol and stained with 0.1% crystal violet to count cell numbers.
7
Cellular Migration and Invasion Assays
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Wound healing and transwell assays were performed to detect the migration and invasion ability. For wound healing assay, transfected cells were inoculated in a 6-well plate. A straight scratch was made in each well when the cell density was close to overgrown, and pictures were taken at the same position at 0 h, 12 h, and 24 h using a microscope. For transwell assay, 1 × 105 cells after 24 h of transfection were seeded onto the upper compartment of the matrigel-coated chamber (Corning Costar, Corning, NY, USA) with 200 μL of serum-free RPMI-1640 medium; in the lower chamber, 600 μL of the medium containing 10% fetal bovine serum were added. Cells on the upper surface were wiped off after 24 h of incubation; the 4% paraformaldehyde was used to fix the invasive cells on the lower surface of the chamber and stained with crystal violet solution.
8
Cell Migration and Invasion Assay
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After 48 h transfection, cells were resuspended into serum-free medium. For migration assays, 5.0 × 104 cells were placed in the top chamber of each insert (Corning Costar, USA) with 8.0 μm pores; for invasion assays, 1.0 × 105 cells were seeded in a Matrigel-coated chamber (Corning Costar, USA). In the lower chamber, 600 μl of RPMI 1640 with 10% FBS was added as a chemoattractant. After the cells were incubated for 24 h, the insert was washed with PBS, and cells on the upper surface of the membrane were removed with a cotton swab. Cells adhering to the lower surface were fixed with methanol, stained with Giemsa. The number of cells in the membrane was counted from 5 randomly selected visual fields with a microscope at 100× magnification. All assays were independently repeated at least three times.
9
Cell Migration and Invasion Assay
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Cell migration assay was implemented using 24-Transwell chambers (8 μm pore size, Costar, Corning, Flintshire, UK) and Matrigel-coated chambers (Costar) were used for cell invasion assay, as described previously.11 (link) To be brief, transfected cells (5.0 × 104) in serum-free media were added into the upper chamber, and the lower chamber was placed with 700 μL of growth media containing 10% FBS as a chemoattractant. Following the 24 h incubation, the images were obtained at ×200 magnification using an Olympus BX41 microscope (Olympus, Tokyo, Japan) and the number of cells that had migrated or invaded through the pores was counted by 10 random fields.
10
Cell Invasion Assay Protocol
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Cell invasion assays were performed in Matrigel-coated chambers (Corning Costar, Corning, NY, USA) with 8-μm pore membranes. The 1 × 105 transfected cells suspended in 200 μL of serum-free RPMI 1640 medium were added to the upper chamber while the lower compartment of the chamber was filled with 600 μL of the medium containing 10% FBS. After 24 h of incubation at 37 °C, the non-migrating cells in the upper chamber were removed by a cotton swab and lower surface of the chamber was fixed with 4% paraformaldehyde and stained with crystal violet. Invading cells were scored by counting at least 5 fields per membrane under a light microscope.
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