Cell invasive or migration capacities were assessed using the BD BioCoat Tumor Invasion System, 24 Multiwell or the BD Falcon FluoroBlok 24 Multiwell Insert System (BD Bioscience) according to the manufacturer’s instructions as described previously [40 (link)]. In brief, miR-203-transfected cells and mock-transfected cells (1.0 x 105 cells/well) were placed in the upper chamber, and the lower chamber was filled with 750 μL of RPMI 1640 with 10% FBS as a chemoattractant, and incubated in a humidified atmosphere (37°C and 5% CO2). After a 48 h incubation, the upper chamber was transferred into a second 24-well plate containing 500 μL of 4 μg/mL calcein AM in HBSS in each well, and the plates were incubated for an additional 1 h (37°C and 5% CO2). Invasive cells that migrated through the membrane were evaluated in a fluorescence plate reader at excitation/emission wavelengths of 485/535 nm. Invasiveness was measured as the percentage of fluorescence of an invasive fibrosarcoma cell line (HT-1080) that served as a control. Each independent experiment was performed three times.
Falcon fluoroblok 24 multiwell insert system
Manufactured by BD
Sourced in United States
The BD Falcon FluoroBlok 24 Multiwell Insert System is a cell culture insert system designed for fluorescence-based applications. It features a 24-well plate format and a black, opaque polyethylene terephthalate (PET) membrane with a pore size of 3.0 μm.
Automatically generated - may contain errors
Lab products found in correlation
Biocoat tumor invasion system, by BD (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Matrigel coating, by BD (1 mentions)
Versamax tunable microplate reader, by Molecular Devices (1 mentions)
Cell proliferation kit 1, by Roche (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
7 protocols using falcon fluoroblok 24 multiwell insert system
1
Quantifying Cell Invasion and Migration
2
Neutrophil Migration Assay under TLR Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PMNs were left untreated or were treated with 10 ng/ml lipopolysaccharide (LPS) or 100 ng/ml Pam3CSK4 for 3, 6 and 12 h in RPMI supplemented with 0.5% FCS. Supernatants were collected and added to the lower well of a BD Falcon™ FluoroBlok™ 24-Multiwell Insert System (BD Biosciences). Additional PMNs were stained for 30 min with 1 µM calcein-AM in HBSS (−Ca2+, −Mg2+) supplemented with 0.5% BSA at 37°C, twice washed and added to the insert (in RPMI supplemented with 0.5% FCS). As positive control of PMN migration 10 nM N-formyl-methionyl-leucyl-phenylalanine (fMLP) was used and a 100% migration control was included. 100 ng/ml Pam3CSK4 or 10 ng/ml LPS in culture medium were used in lower well to exclude PMN migration in response to direct TLR-ligand contact. Migration was measured as fluorescence with a fluorescence microplate reader using excitation at 485 nm and emission detection at 520 nm every second minute over a two hour period at 37°C and 5% CO2.
3
Quantifying Cell Migration and Invasion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell migration was assessed using the BD Falcon FluoroBlok 24 Multiwell Insert System (BD Bioscience, San Jose, CA). The cells (1.0x105 cells/500 μl/well) were then placed in the upper chamber of the 24-well plate with serum-free medium. The lower chamber was filled with 10% fetal bovine serum, which acts as a chemoattractant, and incubated in a humidified atmosphere (37°C and 5% CO2). After a 48 h incubation, invasive cells that migrated through the membrane were evaluated in a fluorescence plate reader at excition /emission wavelengths of 485/530 nm. Invasiveness was measured as the percentage of fluorescence of an invasive fibrosarcoma cell line (HT-1080) that served as a control. Each independent experiment was performed with at least three replicates.
4
Evaluating Cell Invasiveness using Matrigel Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The
invasion assays
were performed as described elsewhere.31 In brief, the direct invasiveness of the cells was evaluated with
the BD Falcon FluoroBlok 24-multiwell insert system (BD Biosciences)
precoated with Matrigel. A migration control was obtained on the same
system without Matrigel coating (BD Biosciences). The top compartments
of both systems was loaded with 60 000 cells per well in minimal
medium (RPMI), and the lower compartment was filled with RPMI with
10% FBS (serving as a chemoattractant). Either aptamer TOV6 or 17-AAG
was added to the minimal medium, which was prefiltered with 0.2 μm
syringe filters (Fisher). The cells were allowed to migrate or invade
by incubating the plates overnight in the cell incubator. The cells
were labeled with calcein AM (Invitrogen) after the invasion or migration
step by injecting the chemical into the lower trans well compartment.
(The plates contain a filter that only allows detection of the cells
in the lower compartment.) Migrated or invaded cells were read on
a VERSAmax tunable microplate reader (Molecular Devices).
invasion assays
were performed as described elsewhere.31 In brief, the direct invasiveness of the cells was evaluated with
the BD Falcon FluoroBlok 24-multiwell insert system (BD Biosciences)
precoated with Matrigel. A migration control was obtained on the same
system without Matrigel coating (BD Biosciences). The top compartments
of both systems was loaded with 60 000 cells per well in minimal
medium (RPMI), and the lower compartment was filled with RPMI with
10% FBS (serving as a chemoattractant). Either aptamer TOV6 or 17-AAG
was added to the minimal medium, which was prefiltered with 0.2 μm
syringe filters (Fisher). The cells were allowed to migrate or invade
by incubating the plates overnight in the cell incubator. The cells
were labeled with calcein AM (Invitrogen) after the invasion or migration
step by injecting the chemical into the lower trans well compartment.
(The plates contain a filter that only allows detection of the cells
in the lower compartment.) Migrated or invaded cells were read on
a VERSAmax tunable microplate reader (Molecular Devices).
5
Cell Proliferation and Migration Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded at 3,000 cells per well in triplicate 96-wells coated with or without collagen I in 100 μl medium, and cell proliferation was evaluated by performing MTT assays using a Cell Proliferation Kit 1 (Roche Applied Science, Penzberg, Germany) according to the manufacturer’s instructions.
Cell-migration capacity was assessed using the BD Falcon FluoroBlok 24-Multiwell Insert System (BD Bioscience, San Jose, CA). Briefly, cells were seeded in the upper chamber of the 24-well plate in serum-free medium. The lower chamber was filled with 750 μl of medium containing 10% FBS, which acts as a chemoattractant, and incubated in a humidified atmosphere (37 °C and 5% CO2). After a 48-hour incubation, the upper chamber was transferred into a second 24-well plate containing 500 μl/well (4 μg/ml) calcein AM in HBSS and incubated for an additional hour (37 °C and 5% CO2). Invasive cells that migrated through the membrane were evaluated in a fluorescence plate reader at excitation and emission wavelengths of 485 and 535 nm, respectively. Three independent experiments were performed. Similarly, cell invasion capacities were assessed using the BD BioCoat Tumor Invasion System, 24 Multiwell (BD Bioscience), following the same procedure used for the migration assays.
Cell-migration capacity was assessed using the BD Falcon FluoroBlok 24-Multiwell Insert System (BD Bioscience, San Jose, CA). Briefly, cells were seeded in the upper chamber of the 24-well plate in serum-free medium. The lower chamber was filled with 750 μl of medium containing 10% FBS, which acts as a chemoattractant, and incubated in a humidified atmosphere (37 °C and 5% CO2). After a 48-hour incubation, the upper chamber was transferred into a second 24-well plate containing 500 μl/well (4 μg/ml) calcein AM in HBSS and incubated for an additional hour (37 °C and 5% CO2). Invasive cells that migrated through the membrane were evaluated in a fluorescence plate reader at excitation and emission wavelengths of 485 and 535 nm, respectively. Three independent experiments were performed. Similarly, cell invasion capacities were assessed using the BD BioCoat Tumor Invasion System, 24 Multiwell (BD Bioscience), following the same procedure used for the migration assays.
6
Evaluating Invasive Capacity of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The BD Falcon FluoroBlok 24 Multiwell Insert System (BD Bioscience, San Jose, CA) was used to evaluate invasive capacity. Twenty-four hours before the assay, cells were transfected with GRB7- and ERBB2-specific siRNAs or a negative control siRNA. The cells (1.0 × 105 cells/500 μL/well) were then placed in the upper chamber of the 24-well plate with serum-free medium. The lower chamber was filled with 750 μL of medium containing 10% FBS, which acted as a chemo-attractant, and plates were incubated in a humidified atmosphere (37°C and 5% CO2). After a 72-h incubation, the upper chamber was transferred into a second 24-well plate containing 500 μL/well calcein AM (4 μg/mL) in Hank’s balanced salt solution (HBSS) and incubated for an additional 1 h (37°C and 5% CO2). Invasive cells that migrated through the membrane were evaluated using a fluorescence plate reader with excitation/emission wavelengths of 485/535 nm. Migration was measured as the percentage of fluorescence emission compared to that of an invasive fibrosarcoma cell line (HT-1080), which served as a control. Each independent experiment was performed 3 times.
7
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 was assessed using The BD Falcon™ FluoroBlok™ 24 Multiwell Insert system (BD Bioscience, San Jose, CA, USA). For a period of 24 h prior to the assay, cells were transfected with PLS3 siRNA or a negative control siRNA. The NUGC3 and AGS cells (1.0×104) were placed in the upper chamber of a 24-well plate with serum-free RPMI-1640 medium. The lower chamber was filled with 750 µl medium with 10% foetal bovine serum, in which the serum acted as a chemoattractant, and the cell migration plate was incubated in a humidified atmosphere (37°C and 5% CO2). After a 48 h incubation, the upper chamber was transferred into a second 24-well plate containing 500 µl/well of 4 µg/ml calcein AM in HBSS (Invitrogen; Thermo Fisher Scientific, Inc.) and incubated for 1 h (37°C and 5% CO2). Invasive cells that migrated through the membrane were evaluated using a fluorescence plate reader at excitation/emission wavelengths of 485/535 nm. Each independent experiment was performed three times. NUGC3 cell invasion was assessed using the BD BioCoat™ Tumor Invasion system 24-Multiwell (BD Bioscience, San Jose, CA, USA) and the same procedure as that used for migration.
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!