The cell invasion assay was performed using 24‐well Corning® BioCoat™ Matrigel Invasion Chambers (Corning). PC cells were seeded into the upper chamber in FBS‐free medium and the lower chamber was filled with medium supplemented with 3% FBS containing M2BPGi at concentrations ranging from 0 to 3 μg/mL. After incubation for 48 h, the cells were fixed and stained with Diff‐Quik (Sysmex). After staining, the cells that migrated through the pores to the lower surface of the membrane were counted under a microscope. In total, five randomly selected fields were evaluated.
Biocoat matrigel invasion chamber
Manufactured by Corning
Sourced in United States, United Kingdom, Austria, Germany
BioCoat Matrigel Invasion Chambers are a laboratory product designed for cell invasion assays. They consist of a cell culture insert with a porous membrane coated with a basement membrane matrix. The chambers allow for the evaluation of a cell's ability to invade through the matrix, which is a key step in many biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Transwell chamber, by Corning (17 mentions)
Diff quik, by Sysmex (15 mentions)
Crystal violet, by Merck Group (10 mentions)
Dmi1 inverted microscope, by Leica (7 mentions)
Diff quick, by Sysmex (7 mentions)
Matrigel, by Corning (6 mentions)
Biocoat control inserts, by Corning (5 mentions)
Cell culture insert, by Corning (5 mentions)
Diff quik stain, by Sysmex (4 mentions)
24 well plate, by Corning (4 mentions)
Diff quik kit, by Sysmex (4 mentions)
Biocoat cell culture chamber, by Corning (3 mentions)
Matrigel, by BD (3 mentions)
Transwell clear polyester membrane inserts, by Corning (3 mentions)
Transwell cell migration assay kits, by Corning (3 mentions)
Bx50 microscope, by Olympus (3 mentions)
Annexin 5 fitc apoptosis detection kit, by Abcam (3 mentions)
Celltiter 96 aqueous one solution cell proliferation assay kit, by Promega (3 mentions)
Falcon cell culture insert, by Corning (3 mentions)
Xtt assay kit 2, by Merck Group (2 mentions)
427 protocols using biocoat matrigel invasion chamber
1
Matrigel Invasion Assay for PC Cell Motility
2
Cell Migration and Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell migration and invasion activity assays were performed as previously described [19 (link)] using Falcon® Permeable Support for-24 Well Plate (8.0 μm pore size) and Corning® BioCoat™ Matrigel® Invasion Chambers (8.0 μm) (Corning, USA). The cells that migrated were stained with a Diff-Quik Kit (Medion Diagnostics, Switzerland) and counted at a 40x magnification under the Olympus BX41 microscope. Each experiment was performed in triplicate and repeated three times.
3
Invasive Potential of TAM-Conditioned HSC-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HSC-2 cell lines were cultured for 4 days in CM of CD163+, CD204+, and CD206+ cells and used for invasion assays by using Corning® BioCoat™ Matrigel® Invasion Chambers (Corning Incorporated, Corning, NY, USA) As follows the product’s guidelines, three Matrigel and three control inserts were prepared for a 24 well culture dish. The CM (750 μl) of CD163+, CD204+, and CD206+ TAM was used as a chemoattractant in each well subsequently and the insert was filled with 500 μl of DMEM (Life Technologies Corporation) without serum and HSC-2 cell line (2.5 × 104 cells/insert). The cells were cultured for 22 hours in a humid chamber with 5% CO2 atmospheric state at 37 °C temp. Then the inserts were removed from the wells and Matrigel was cleaned off. After that cells were fixed with 100% methanol (Junsei Chemical Co.,Ltd., Tokyo, Japan) and then stained with Mayer’s hemalum solution (Merck KGaA, Darmstadt, Germany, 1:4 dilution) and Tissue-Tek eosin (Sakura Finetek Japan Co., Ltd, Tokyo, Japan) (Supplementary Method 1 ).
The numbers of cells on the membrane were counted in 4 mm2 sections from five different high-power microscopic fields (400×; 0.0625 μm2). The average number of cells in the Matrigel insert membrane was divided with the average number of cells in the control insert membrane, and then multiplied with 100 to calculate the invasion percentage.
The numbers of cells on the membrane were counted in 4 mm2 sections from five different high-power microscopic fields (400×; 0.0625 μm2). The average number of cells in the Matrigel insert membrane was divided with the average number of cells in the control insert membrane, and then multiplied with 100 to calculate the invasion percentage.
4
Quantifying Cell Invasion via Matrigel Chambers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 6-well plates and transfected with 10 nM of htRNALeu/miR-34a-5p, htRNALeu/miR-124-3p or htRNALeu. After 48 h, survived cells were harvested and seeded onto the upper inserts of the 24-well Corning BioCoat Matrigel Invasion Chambers (Corning, Bedford, MA) at 6 × 104 cells/well with 500 μL serum-free RPMI 1640 medium where the lower chamber was supplied with 750 μL of serum-containing culture medium (10% FBS). 20 h later, the invaded cells were fixed with 500 μL of 10% formaldehyde, stained with 500 μL 0.1% crystal violet, and imaged with an Olympus IX2-UCB microscope. The number of invaded cells was acquired by counting five fields per insert (100 × magnification), and then compared between different treatments as we described recently 31 (link).
5
Cell Migration and Invasion Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell migration and invasion chamber assays were performed with the Corning Transwell migration chambers and Corning BioCoat Matrigel Invasion chambers (Corning Inc., Corning, NY, USA) according to the manufacturer’s instructions. Briefly, 1×105 cells were seeded into the upper chamber in serum-free medium. After incubation for 24 hours, nonmigrating or invading cells on the surface of the upper chamber were removed with a cotton-tipped swab. The migrated or invaded cells on the lower side were fixed and stained. The cells were photographed under a microscope and counted with ImageJ software. Each experiment was repeated for three times.
6
Cell Invasion Assay with Matrigel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell invasion assays were performed using 24-well Corning BioCoat™ Matrigel Invasion Chambers (Corning, New York, NY, USA) according to the manufacturer’s protocol. Five hundred microlitres of medium containing 10% FBS and MIC-1 were added to the lower chamber, while HuCCT-1 and TFK-1 cells (2.0 × 105 cells/well) were added to the upper chamber in 500 µl of serum-free medium. After 22 h of incubation, cells that did not invade through the membrane were removed from the upper chamber using a cotton swab. The invaded cells were fixed with 4% paraformaldehyde for 20 min and stained with a 1% crystal violet solution for 30 min at room temperature. Three different fields were photographed using a BX41-13 microscope (Olympus, Tokyo, Japan) at 200 × magnification, and invasive cells were counted.
7
Cell Migration and Invasion Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Migration assays were performed in cell culture inserts (24-well, 8-µm pore size, #353097, Corning, Inc.). Cell concentrations ranged from 105 to 2×105 cells/ml. Invasion assays were also performed in Corning BioCoat Matrigel invasion chambers (24-well, 8-µm pore size, #353097, Corning, Inc.). Cell concentrations ranged from 2×105 to 4×105 cells/ml. Cells were seeded on uncoated or Matrigel-coated inserts in 500 ml of serum-free medium for migration and invasion assays, respectively. The lower chambers were filled with 750 µl of 10% FBS-supplemented medium. After 48 h, the cells on the lower surface of the insert were fixed and stained with crystal violet (Differential Quik Stain kit, Polysciences) at room temperature for 2 min. The number of stained cells was counted in >3 fields under an inverted microscope (Olympus CKX31).
8
Functional Assays for Head and Neck Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The procedures for conducting functional assays (cell proliferation, migration, and invasion assays) with HNSCC cells were described in earlier publications [23 (link),24 (link),25 (link),26 (link)]. In brief, for proliferation assays, Sa3 or SAS cells were transferred to 96-well plates at 3.0 × 103 cells per well. Cell proliferation was evaluated using the XTT assay kit II (Sigma–Aldrich, St. Louis, MO, USA) 72 h after the transfection procedure. For migration and invasion assays, Sa3 and SAS cells were transfected in 6-well plates at 2.0 × 105 cells per well. After 48 h, transfected Sa3 and SAS cells were added into each chamber at 1.0 × 105 per well. Corning BioCoatTM cell culture chambers (Corning, Corning, NY, USA) were used for migration assays whereas Corning BioCoat Matrigel Invasion Chambers were used for invasion assays. After 48 h, the cells on the lower surface of chamber membranes were stained and counted for analysis. All experiments were performed in triplicate.
9
Cell Invasion Assay Using Matrigel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell-invasion assay was performed using 24-well Corning® BioCoat™ Matrigel Invasion Chambers (Corning, NY, USA). HCC cell lines in FBS-free medium were seeded into the Matrigel inserts at a density of 1 × 105 per chamber. The lower chamber contained 1 or 3 µg/ml M2BPGi in medium containing 10% FBS. After incubation for 48 h, the cells were fixed and stained with Diff-Quik (Sysmex Corporation, Kobe, Japan). After staining, cells that migrated through the pores to the lower surface of the membrane were counted under the microscope. In total, five randomly selected fields were evaluated.
10
Evaluating miRNA and KRT80 Functions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tumor-suppressive functions of miRNAs were evaluated by transient transfection assays using mature miR-139-5p and miR-139-3p. The tumor-promoting functions of KRT80 (loss-of-function assays) were assessed by siRNA transfection assays using siRNAs targeting KRT80. Functional assays (proliferation, migration, and invasion assays) were performed according to procedures of previous studies [51 (link),52 (link)]. Briefly, for proliferation assays, HCT116 or DLD-1 cells were transferred into 96-well plates at 3.0 × 103 cells/well. Cell proliferation was assessed using XTT assay kit II (Sigma-Aldrich, St. Louis, MO, USA) 72 h after the transfection procedure. For the migration and invasion assay, HCT116 and DLD-1 cells were transfected in 6-well plates at 3.0 × 105 cells/well; 48 h later, transfected HCT116 and DLD-1 cells were added to each chamber at 1.0 × 105 cells/well. Corning BioCoatTM cell culture chambers (Corning, Corning, NY, USA) were used for the migration assay and Corning BioCoat Matrigel Invasion Chambers were used for the invasion assay. cells on the underside of the chamber membrane were stained and counted for analysis. All experiments were performed in triplicate. The details of the reagents used in these analyses are listed in Table S2 .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!