HEK293, RAMOS and Jurkat human cell lines were obtained from ATCC. Cells were cultured in RPMI supplemented with 10% fetal calf serum and pen/strep. Cells were passaged and cultured at 37° C 5% CO2 atmosphere. Cell viability and counting was performed on Countess II automated cell counter (Invitrogen). RAMOS and Jurkat cells were cultured between 0.2 and 2.0e6 cells/ml. HEK cells were passaged 2–3 times per week with warm trypsin (Invitrogen). Cell proliferation was measured by electrical impedance using an xCelligence RTCA DP analyzer (Agilent) with E-plate 16 glass plates. HEK293 cells were harvested and counted and further diluted into E-plate 16 wells in 100 μL media. Cys-S3 or Cys-SS− was added (1:100 dilution) to wells and plates were loaded into the analyzer in a humidified, 5% CO2 incubator at 37° C. Cell index measurements were recorded every 15 min and normalized to wells containing media without cells.
Xcelligence rtca dp analyzer
Manufactured by Agilent Technologies
Sourced in United States
The XCELLigence RTCA DP Analyzer is a real-time cell analysis system used for monitoring cellular events in a label-free and non-invasive manner. The device measures changes in electrical impedance of cells adhering to microelectronic sensor arrays, providing quantitative information on cell number, cell size, and cell attachment or spreading.
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Lab products found in correlation
13 protocols using xcelligence rtca dp analyzer
1
Cell Viability Assay with Cys-S3 and Cys-SS-
2
Allogeneic T Cell Cytotoxicity Assay
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All experiments were performed with T cells isolated from healthy donors as described above. Isolated T cells were primed with fully HLA-expressing LSCs which were previously stimulated with IFNɣ (100ng/mL) for 48 hours. T cell priming was performed for 7 days in RPMI 1640 Medium (Lonza, Basel, Switzerland) supplemented with 5% AB serum and IL-2 (100U/ml) (Prepotech, New Jersey, USA), IL-7 (100ng/mL) (Prepotech, New Jersey, USA) and IL-12 (50ng/mL) (Prepotech, New Jersey, USA). Real time cytotoxicity was measured using XCelligence RTCA DP analyzer (Agilent Technologies, California, USA). First, background signal was measured using E-plate 16 (Agilent Technologies, California, USA) with 50µl medium. Silenced and non-silenced LSCs (target cells) were seeded and let adhere for 24 hours. Subsequently, pre-stimulated allogeneic T cells (effector cells) were added in a 1:2 ratio (target: effector). Changes in electrical impedance were expressed as cell index values, which correlates with cellular coverage of electrode sensors at the bottom of each well, and normalized to baseline impedance values before adding effector cells.
3
Real-Time Cell Proliferation Assay
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Serial 15-min interval real-time proliferation measurement of cells was performed for 48 h using xCELLigence RTCA DP analyzer (Agilent Technologies) in a cell culture incubator (37 C, 5% CO 2 ).
Using a 16-well electronic microtiter plate (16-PET plate), technical replicates of 2 Â 10 4 cells were seeded per condition.
Following the manufacturer's instructions, background measurement was performed using 50 μl of culture medium, then cells were added to a final volume of 100 μl and incubated for 30 min until fully attached.
Using a 16-well electronic microtiter plate (16-PET plate), technical replicates of 2 Â 10 4 cells were seeded per condition.
Following the manufacturer's instructions, background measurement was performed using 50 μl of culture medium, then cells were added to a final volume of 100 μl and incubated for 30 min until fully attached.
4
Cell proliferation and migration analysis
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Cell proliferation and migration were analyzed with RTCA assay as described before (32 ). Cells were cultured at 6000 per well in CIM-Plate wells coated with (invasion) or without (proliferation) matrigel. The cell index signals were read by xCELLigence RTCA DP Analyzer (ACEA Bioscience). Invasion and migration are monitored continuously over a 48-hour period. Each experiment was repeated three times and results were presented as mean ± SD.
5
CD99-Mediated Cell Migration Assay
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HeLa cells were transfected with different CD99 constructs, detached using a cell scraper, counted, and resuspended in DMEM, 1% FCS. The real-time migration assay was performed using the xCELLigence RTCA DP analyzer (ACEA Biosciences). A CIM-Plate 16 was assembled and equilibrated with DMEM, 1% FCS for 1 h. Then 40,000 cells were added and allowed to migrate through the microporous membrane for 24 h. Results are expressed as cell index (CI), a relative unit reflecting the electrical impedance measurement at the bottom of the microporous membrane.
6
Real-Time Cell Proliferation Analysis
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Cell proliferation was analyzed with RTCA instrument (ACEA Bioscience Inc.) according to the manufacturer’s instruction as described before25 (link). Cells were cultured at 10,000 per well in CIM-Plate wells. The cell index signals were read by xCELLigence RTCA DP Analyzer (ACEA Bioscience Inc.).
7
Cell Proliferation and Migration Assay
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Cell proliferation and migration were analyzed with RTCA assay as described before [39 (link)]. Cells were cultured at 6000 per well in CIM-Plate wells coated with (invasion) or without (migration) matrigel. The cell index signals were read by xCELLigence RTCA DP Analyzer (ACEA bioscience Inc). Invasion and migration are monitored continuously over a 48-hour period. Each experiment was repeated three times and results were presented as mean ± SD.
8
Transwell Migration Assay with RTCA
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Migration through myelin-coated and electronically integrated transwells was monitored using an xCELLigence RTCA DP analyzer (Acea Biosciences, USA).
9
Real-time Analysis of Tumor Cell Dynamics
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MCA tumor cell lines were seeded into E-Plate 16 (ACEA Biosciences, San Diego, CA) once they had reached log phase growth in T-75 cell culture flask. 200 μl of complete RPMI medium (RPMI GlutaMAX, 10% FCS and 1% PenStrep) containing 5x104 cells/ml of cells were transferred to E-Plates. E-Plates were incubated for 30 minutes to allow for settling of cells and then transferred into the xCelligence RTCA DP Analyzer (ACEA Bioscience) and cell indexes were recorded every 30 minutes over a 7-day period. Growth rate (slope) was determined using linear regression analysis where 95% R2 has been observed between two time points at the logarithmic growth phase of the cells. Cisplatin (5 μg/ml) was added when cells were determined to enter the log-phase growth. Death rate induced by Cisplatin was determined similarly to the growth rate from the moment effect of cisplatin was observed.
10
Transwell and RTCA Assays for Cell Invasion and Migration
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The RTCA assay was done as the manufacturer’s protocol. Cells were cultured at 6000 per well in CIM-Plate wells coated with (invasion) or without (migration) matrigel. The cell index signals were read by xCELLigence RTCA DP Analyzer (ACEA Bioscience Inc.). Invasion and migration are monitored continuously over a 48-h period.
The transwell assay was done as follows. Briefly, cells were split at 1 × 105 per well in 24-well transwell plates coated with (invasion) or without (migration) matrigel. The cells were fixed in 4 % PFA and stained by crystal violet after 48 h. The positive cells were counted under microscope.
Each experiment was repeated three times and results were presented as mean ± SD.
The transwell assay was done as follows. Briefly, cells were split at 1 × 105 per well in 24-well transwell plates coated with (invasion) or without (migration) matrigel. The cells were fixed in 4 % PFA and stained by crystal violet after 48 h. The positive cells were counted under microscope.
Each experiment was repeated three times and results were presented as mean ± SD.
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