A cell proliferation assay was conducted with the CellTrace proliferation kit (catalog# C34570, Thermo Fisher Scientific) following the protocol provided by the vendor. A375 cells were suspended in DPBS to attain a cell density of 1 × 106 cells per mL. Then, 1 µL of carboxyfluorescein succinimidyl ester (CFSE) dye was added to 1 mL of the cell suspension and the cells were incubated at room temperature for 20 min. After incubation, DMEM medium was added to the cell suspension to stop the staining and the cells were centrifuged at 900 rpm for 5 min. The supernatant was removed, and the pellet was resuspended in fresh DMEM medium. Approximately 3 × 105 of CFSE-stained cells were plated in each well of a 6-well plate and incubated for 24 h. After 24 h, the cells were either treated with 10 µM of VEM or left untreated. At indicated time points, cells were collected, centrifuged, and resuspended in 1 mL of DPBS. The cells were then stained with 1.5 µM of PI to detect dead cells. The cells were transferred to a 5 mL test tube and analyzed with a flow cytometer to measure green (excitation: 488 nm; emission: 520 nm) and red (excitation: 561 nm; emission: 586 nm) fluorescence, respectively.
Celltrace proliferation kit
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
The CellTrace Proliferation Kit is a fluorescent cell labeling solution designed for tracking cell division and proliferation. It provides a simple and effective method to monitor the proliferation of cells in culture.
Automatically generated - may contain errors
Lab products found in correlation
10 protocols using celltrace proliferation kit
1
Cell Proliferation Assay with CFSE
2
CD4+ T Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proliferation of CD4+ T cells purified by negative sorting (Thermo, 11416D) was assessed using the CellTrace Proliferation Kit (Thermo, C34554) according to the manufacturer’s instructions.
3
Cell Proliferation Assay Using CellTrace
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell proliferation assay was performed using CellTrace Proliferation Kit (Thermo Fisher). Cells were stained according to the manufacturer's protocol, and the mean fluorescence intensity (MFI) was measured. Further readings were followed daily. MFI values were used to calculate the proliferation index (43 (link)). Details are in SI Appendix, Methods .
4
Evaluating T-cell Proliferation and Cytokine Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice splenocytes were obtained as described previously (5 (link)). Both primary skin fibroblast-like cells (104) and SphCs (104) were cocultured for 72 h with 5×105 splenocytes activated with anti-CD3 antibody (1 mg/mL, eBioscience). A CellTrace Proliferation Kit (Thermo Fisher Scientific, USA) was used to evaluate T-CD4+ proliferation by staining with anti-CD4-APC. Cell media were harvested at 36 h, and cytokines were analyzed by FACS using a Cytometric Bead Array Mouse Th1/Th2/Th17 Cytokine Kit (BD Biosciences).
5
Cell Proliferation Assay with CellTrace
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell proliferation assay was performed using CellTrace Proliferation Kit (ThermoFisher) according to the manufacturer's protocol. In brief, one million cells were washed with PBS and stained in a 1/1000 dilution of the stock solution for 20 min at 37°C protected from light. Cells were washed twice with complete supplemented RPMI. After 10 min the mean fluorescence intensity (MFI) was measured. Further readings were followed daily. MFI values were used to calculate the proliferation index 37 . Proliferation was measured at least three times.
6
Cell Proliferation Assay in MEC-1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proliferation was evaluated in NICD-transfected MEC-1 cells by using CellTrace Proliferation Kit (for flow cytometry; ThermoFisher Scientific), as reported previously. 41 Further details regarding the materials and methods used are provided as Supplementary Information.
7
Quantifying Cell Proliferation Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine cell proliferation, cells over-expressing either MeCP2-WT or K171Q mutant were stained with carboxyfluorescein succinimidyl ester (CFSE) from Cell Trace Proliferation kit from Invitrogen. Cells were stained according to manufacturer's guidelines. 1×106 cells/ml were incubated in a suspension with 0.1% BSA in PBS. Cells were incubated with 10μM of CFSE for 10 minutes at 37°C. Residual stain was removed from cells and cells were placed back at 37°C for 72 hours. Cells were harvested using 1mM EDTA, and then resuspended in FACS buffer (PBS, 2% FBS, 0.1% sodium azide). Cells were analyzed using FACS Calibur.
8
Antigen-Specific T Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4+ and CD8+ T cells were electroporated with RNA coding for the gp100-specific TCR and were subsequently labelled with CFSE according to the Cell Trace proliferation kit (Invitrogen, ThermoFisher Scientific, Darmstadt, Germany) using anhydrous DMSO. Co-cultures between T cells and peptide-loaded or non-peptide-loaded moDCs was performed as described above at a 1:1 ratio. BRAF and/or MEK inhibitors were applied at the indicated concentration (Table 1 ). After three days, the cells were harvested and were analyzed using flow cytometry. To assess antigen-specific vs. the spontaneous proliferation, we calculated the percentage of proliferated CD8+ or CD4+ T cells either after antigen-specific (gp100 peptide) or unspecific (no peptide) stimulation, respectively.
9
Isolation and Activation of Mouse CD4+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD4+ T cells were isolated from spleens of mice with a magnetic microbeads negative selection kit (EasySep Mouse CD4+ T Cell Isolation Kit; STEMCELL Technologies) and labeled with 5 μM carboxyfluorescein diacetate succinimidyl ester (CFSE; CellTrace Proliferation Kit, Invitrogen) for 5 min in PBS containing 5% fetal bovine serum, at room temperature. After incubation, cells were washed twice with PBS, resuspended in RPMI-enriched medium (composition stated above), and activated with anti-CD3/anti-CD28 beads (Dynabeads, Gibco) in 96-well (U shape) plates (80 × 105 cells per well). After 72 hours, cells were collected and analyzed by flow cytometry.
10
In Vitro Suppression Assay of Tregs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For in vitro suppression assay, naïve CD4+ T cells were isolated from spleens of young (2 months) CD45.1 mice using a naïve isolation kit (EasySep Mouse Naïve CD4+ T Cell Isolation Kit, STEMCELL Technologies), labeled with CFSE (CellTrace Proliferation Kit, Invitrogen), and used as responder cells (2 × 104 cells per well). Then, cells were cultured in 96-well plates with irradiated 2 × 104 APCs (as feeder cells) in the presence of sorted CD25highCD81− or CD25highCD81+ Tregs at 1:1, 1:2, and 1:4 responders:Tregs ratios. Cells were stimulated with anti-CD3 (1 μg/ml) for 72 hours. Proliferation (defined as all cells with CFSE dilution) of responder cells was analyzed to assess the suppression of Tregs cells. The percentage of suppression was determined as [100 − (% of proliferating cells with Tregs)/(% of proliferating cells without Tregs)].
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!