Invasion assays were performed using chamber with 8 μm pore filters (Corning, New York, NY) pre-coated with 1 mg/mL Matrigel (BD Biosciences, Shanghai, China). Briefly, CNE-1 cells and HONE-1 cells were transfected with siRNA1 or siNC. At 24h post transfection, the cells were incubated in serum-free medium overnight. The cells were collected and exact 5 × 104 cells in serum-free medium were plated to the upper chamber, while medium containing 10% FBS was added to the lower chamber. After 24h of incubation, cells on the lower surface of the membrane were fixed with 4% paraformaldehyde, stained with 0.5% crystal violet and then counted in five random fields. To block cell proliferation, mitomycin C (5 μg/mL, Sigma, Shanghai, China) was added.
Chamber with 8 μm pore filters
Manufactured by Corning
Sourced in United States
The chamber with 8 μm pore filters is a laboratory equipment designed for filtration processes. It features eight apertures, each containing a filter with 8 micron pore size. The core function of this product is to facilitate the separation of particles or substances from a liquid or gas sample by passing it through the porous filters.
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Lab products found in correlation
3 protocols using chamber with 8 μm pore filters
1
Invasion Assay for Metastatic Potential
2
TRIM6 Modulates Cell Migration
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Transwell assay was performed using chamber with 8 μm pore filters (Corning, New York, NY, USA). HK2 cells were transduced with virus expressing TRIM6 shRNAs (shTRIM6-1#, 2#) or the control shRNA (shNC). After 24 h, the cells were trypsinized and plated onto the upper chamber. 1 μM of Ang II was added to the lower chamber. After incubation for 24 h at 37°C, non-migrating cells were completely removed, and the migrated cells were fixed in 4% paraformaldehyde, stained with 0.5% crystal violet, visualized under a microscope, and counted in five random fields.
3
Cell Migration and Invasion Assay
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Migration of cells was assayed using chamber with 8 μm pore filters (Corning, New York, NY, USA). Cells were grown to about 50% confluency and transfected with the desired siRNA. After 24 h, the cells were incubated in serum-free medium for 24 h. Then cells were trypsinized and 5 × 104 cells in serum-free medium added to the upper chamber. Then media with 10% FBS was added to the lower chamber. Cells were incubated for 24 h at 37°C, and then non-migrating cells were completely removed. Cells that migrated to the bottom of the membrane were then fixed in 4% paraformaldehyde and stained by 0.5% crystal violet. Then stained cells were visualized under a microscope, counted in five random fields, and the average number was taken.
For invasion assay, the upper chamber was pre-coated with 1 mg/ml Matrigel (BD Biosciences). The rest of the assay was performed as migration assay.
For invasion assay, the upper chamber was pre-coated with 1 mg/ml Matrigel (BD Biosciences). The rest of the assay was performed as migration assay.
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