Transwell assay was performed using a 24-well plate inserts with 8 μm pores (Corning Incorporated, Corning, NY, USA) to evaluate cell invasion capacity. GI-LI-N and SK-N-BE(2) cells (2×104 cells/well) re-suspended in DMEM medium alone (100 μL) were placed into the top chamber pre-coated with Matrigel (BD Bioscience, Franklin Lakes, NJ, USA). To induce cells invading through the membrane, the bottom chamber was filled with DMEM containing 10% FBS. After 24 h of incubation, non-invaded cells were carefully removed with cotton bud, and invaded cells were fixed using ethanol (95%) and then stained using crystal violet (0.1%). Finally, a microscope (Olympus, Tokyo, Japan) was applied to observe and count invaded cells in five random fields.
24 well plate inserts with 8 μm pores
Manufactured by Corning
Sourced in United States
The 24-well plate inserts with 8 μm pores are a laboratory equipment product designed for cell culture applications. The inserts feature a porous membrane with a pore size of 8 μm, allowing for the study of cell migration, invasion, and other cellular processes.
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4 protocols using 24 well plate inserts with 8 μm pores
1
Transwell Cell Invasion Assay
2
Transwell Invasion Assay for Cell Motility
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Transwell assay was performed using a 24-well plate inserts with 8 μm pores (Corning Incorporated, Corning, NY, USA) to evaluate cell invasion capacity. GI-LI-N and SK-N-BE(2) cells (2×10 4 cells/well) resuspended in DMEM medium alone (100 μL) were seeded into the top chamber pre-coated with Matrigel (BD Bioscience, Franklin Lakes, NJ, USA). To induce cells invading through the membrane, the bottom chamber was lled with DMEM containing 10% FBS (600 μL). After incubation for 24 h, non-invaded cells were carefully removed with a cotton bud, and invaded cells were xed with 95% ethanol and then stained with 0.1% crystal violet. Finally, invaded cells from ve random elds were photographed and counted by a microscope.
3
Transwell Invasion Assay for Cell Motility
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell assay was performed using a 24-well plate inserts with 8 μm pores (Corning Incorporated, Corning, NY, USA) to evaluate cell invasion capacity. GI-LI-N and SK-N-BE(2) cells (2×10 4 cells/well) resuspended in DMEM medium alone (100 μL) were seeded into the top chamber pre-coated with Matrigel (BD Bioscience, Franklin Lakes, NJ, USA). To induce cells invading through the membrane, the bottom chamber was lled with DMEM containing 10% FBS (600 μL). After incubation for 24 h, non-invaded cells were carefully removed with a cotton bud, and invaded cells were xed with 95% ethanol and then stained with 0.1% crystal violet. Finally, invaded cells from ve random elds were photographed and counted by a microscope.
4
Transwell Invasion and Metabolism Assays
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Transwell assay was performed using a 24-well plate inserts with 8 μm pores (Corning Incorporated, Corning, NY, USA) to evaluate cell invasion capacity. GI-LI-N and SK-N-BE(2) cells (2×10 4 cells/well) resuspended in DMEM medium alone (100 μL) were seeded into the top chamber pre-coated with Matrigel (BD Bioscience, Franklin Lakes, NJ, USA). To induce cells invading through the membrane, the bottom chamber was lled with DMEM containing 10% FBS (600 μL). After incubation for 24 h, non-invaded cells were carefully removed with a cotton bud, and invaded cells were xed with 95% ethanol and then stained with 0.1% crystal violet. Finally, invaded cells from ve random elds were photographed and counted by a microscope.
Measurement of glucose consumption and lactate production GI-LI-N and SK-N-BE(2) cells (5×10 4 cells/well) were seeded into six-well plates and transfected with oligonucleotide or/and plasmid. Cell culture media were collected 48 h after the transfection. In accordance with the manufacturer's instructions, the glucose consumption and lactate production were detected with the glucose assay kit (Sigma-Aldrich) and lactate assay kit (BioVision, Mountain View, CA, USA), respectively. A standard calibration curve was used for determining the data, followed by normalizing to the amount of total protein.
Measurement of glucose consumption and lactate production GI-LI-N and SK-N-BE(2) cells (5×10 4 cells/well) were seeded into six-well plates and transfected with oligonucleotide or/and plasmid. Cell culture media were collected 48 h after the transfection. In accordance with the manufacturer's instructions, the glucose consumption and lactate production were detected with the glucose assay kit (Sigma-Aldrich) and lactate assay kit (BioVision, Mountain View, CA, USA), respectively. A standard calibration curve was used for determining the data, followed by normalizing to the amount of total protein.
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