TE1 cell cycle distribution was measured using a flow cytometer by quantifying the PI labeled DNA content. After transfection with recombinant plasmids for 48 h, TE1 cells were collected and fixed with 70% cold ethanol at 4°C for 30 min, then washed with PBS. Pellets were re-suspended and incubated in PBS containing RNaseA (50 µg/ml), Triton (0.2%) and PI (20 µg/ml) at 4°C for 30 min in the dark. Cell cycle analysis was performed using a flow cytometer (BD Calibur; Becton-Dickinson and Company). Data were analyzed with Cell Quest data acquisition and analysis software (Becton-Dickinson and Company) after 1 h.
Cell quest data acquisition and analysis software
Manufactured by BD
Sourced in United States
Cell Quest is a data acquisition and analysis software designed for flow cytometry applications. It provides tools for collecting, organizing, and analyzing data generated from flow cytometers. The software enables users to capture, visualize, and interpret cellular characteristics and populations.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using cell quest data acquisition and analysis software
1
Cell Cycle Analysis by Flow Cytometry
2
Quantification of Apoptosis in TE1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The quantification of apoptotic TE1 cells was performed using flow cytometry with Annexin V-FITC and PI dyes, according to the manufacturer's instructions. A total of 7.5×105 TE1 cells in 6-well plates were transfected with 4 µg recombinant plasmids pcTERT-melittin or pcTERT for 24, 48 and 72 h, before being harvested via trypsinization and washed with PBS. Subsequently, 70% cold ethanol was used to fix the TE1 cells for 30 min at 4°C. Pellets were re-suspended and incubated in Annexin V-FITC labelling solution at 15–25°C for 15 min. PI was added at 4°C and stained for 5 min. The apoptotic cells were then analyzed using a flow cytometer (BD Calibur; Becton-Dickinson and Company). Data were analyzed with Cell Quest data acquisition and analysis software (Becton-Dickinson and Company).
3
Tumor Dissociation and Flow Cytometric Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tumors were weighed, minced into small fragments, and digested in medium containing 0.1 mg/ml DNase (Sigma-Aldrich) and 1 mg/ml collagenase IV (Sigma-Aldrich) at 37°C for 1 h (Zhang et al., 2011 (link); Yang et al., 2017 (link)). The dissociated cells were then prepared for analysis by flow cytometry.
Antibodies targeting CD3ε, CD4, CD8, CD11b, CD80, CD86, CD54, I-a, CD11c conjugated to the corresponding fluorescent dyes were purchased from eBioscience (San Diego, CA, United States). Single-cell suspensions (1 × 106 cells) were stained with different monoclonal antibodies, according to the manufacturer’s instructions. Then, samples were analyzed on a FACSuite using the CellQuest data acquisition and analysis software (BD Biosciences, CA, United States).
Antibodies targeting CD3ε, CD4, CD8, CD11b, CD80, CD86, CD54, I-a, CD11c conjugated to the corresponding fluorescent dyes were purchased from eBioscience (San Diego, CA, United States). Single-cell suspensions (1 × 106 cells) were stained with different monoclonal antibodies, according to the manufacturer’s instructions. Then, samples were analyzed on a FACSuite using the CellQuest data acquisition and analysis software (BD Biosciences, CA, United States).
4
Annexin V-FLUOS Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis of apoptotic cells was conducted using annexin V-fluorescein and PI staining. In accordance with the manufacturer's protocol (Wuhan Hualian Branch Biotechnology Co., Ltd.), transfected or untransfected control MDA-MB-231 cells were collected by trypsinization, washed with PBS, resuspended in 100 µl annexin V FLUOS labeling solution and incubated for 10–15 min at 15–25°C. Cellular apoptosis was evaluated using flow cytometry. Flow cytometric analysis clearly differentiated normal (living) cells, which exhibit low annexin V and low PI staining, from apoptotic cells (high annexin V and low PI staining) and necrotic cells (high annexin V and high PI staining). The data were analyzed by using CellQuest data acquisition and analysis software (version 5.1; BD Biosciences, Franklin Lakes, NJ, USA).
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!