Cell death was analyzed by Annexin-V FITC/propidium iodide (PI) staining (BD Biosciences). Cells were seeded in 6-well plates and transfected with the LINC00365 overexpression plasmid or empty vector. The attached and detached cells were harvested and subjected to Annexin-V FITC/PI staining according to the manufacturer’s instructions. Samples were analyzed using an Accuri C6 Flow Cytometer (BD Biosciences).
Annexin 5 fitc propidium iodide staining
Manufactured by BD
Sourced in United States
Annexin V-FITC/propidium iodide (PI) staining is a laboratory technique used for the detection and analysis of apoptosis, a programmed cell death process. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the outer cell membrane during apoptosis. FITC (fluorescein isothiocyanate) is a fluorescent dye that is conjugated to Annexin V, allowing for the visualization of apoptotic cells. Propidium iodide (PI) is a fluorescent dye that binds to DNA and is used to identify cells with compromised cell membranes, which are typically associated with late apoptosis or necrosis.
Automatically generated - may contain errors
Lab products found in correlation
Accuri c6 flow cytometer, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
Annexin 5 fitc and pi, by BD (1 mentions)
Acea flow cytometer, by Agilent Technologies (1 mentions)
Novoexpress version 1, by Agilent Technologies (1 mentions)
Annexin 5 fitc pi, by Merck Group (1 mentions)
Pi rnase staining solution, by BD (1 mentions)
Annexin 5 pe 7aad assay, by BD (1 mentions)
Navios flow cytometer, by Beckman Coulter (1 mentions)
Flow cytometry, by BD (1 mentions)
7 protocols using annexin 5 fitc propidium iodide staining
1
Annexin-V FITC/PI Cell Death Assay
2
Quantifying Cell Apoptosis via Annexin V-FITC/PI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was performed using Annexin V-FITC/Propidium Iodide (PI) staining (BD PharMingen, San Jose, CA, USA). GC-1 cells were seeded in 6-well plates at a concentration of 106 cells mL-1. The cells were labeled with Annexin V-FITC for 5 min in the dark. Then, 5 mg ml-1 PI was added to each sample for 30 min so that flow cytometry could be performed (BD PharMingen, San Jose, CA, USA).
3
Quantifying Apoptosis by Annexin V-FITC/PI Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was quantitated by flow cytometry after staining cells with annexin V-FITC/propidium iodide (PI) staining (BD). Briefly, after indicated treatments, cells were trypsinized and washed twice with cold PBS, and then resuspended in 1 × binding buffer with 5 μl annexin V and 5 μl of PI at a concentration of 5 × 105/mL cells in a total volume of 100 μl. The cells were gently mixed and incubated in the dark for 15 minutes at room temperature before the number of apoptotic cells was quantified by flow cytometry (LSRFortessa, BD) within 1 hour.
4
Evaluating Cell Death Mechanisms
5
Apoptosis and Mitochondrial Membrane Potential in FaDu Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FaDu cells were transfected with pCMV6-BCAT1, BCAT1 siRNA or corresponding negative controls. At 48 h later, the cells were treated with 2 µM cisplatin (cat. no. P4394; Sigma-Aldrich; Merck KGaA) at 37°C for 24 h. Then the cells were stained with Annexin V-FITC/PI or JC-1 respectively. The rate of apoptosis was determined using Annexin V-FITC/propidium iodide (PI) staining (BD Biosciences). Cells were incubated with 5 µl FITC Annexin V and 5 µl PI for 15 min at room temperature in the dark. The apoptotic rate was calculated as the percentage of early + late apoptotic cells. JC-1 staining was used to determine the mitochondrial membrane potential (Δψm). The cells were harvested, washed with PBS and incubated with 20 µM JC-1 (cat. no. ab113850; Abcam) for 10 min at room temperature. The cells were analyzed using an ACEA flow cytometer (ACEA Bioscience, Inc.; Agilent Technologies, Inc.). Data were analyzed using NovoExpress version 1.2.4 software (ACEA Bioscience, Inc.; Agilent Technologies, Inc.).
6
Apoptosis and Cell Cycle Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Both cell apoptosis assays and cell cycle assays were performed on Navios flow cytometer (Beckman Coulter, CA, USA). Cell apoptosis was detected by Annexin V-PE/7-AAD assay or Annexin V-FITC/ propidium iodide (PI) staining according to the manufacturer’s instructions (BD Biosciences, MA, USA). For cell cycle analysis, cells were washed with PBS, fixed with 70% ethanol overnight at 4 °C, and resuspended in PI/RNase staining solution (BD Biosciences). The percentage of cells in the indicated cell cycle phase was calculated with ModFit LT version 3.2 software.
7
Annexin V-FITC/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The apoptotic cells were detected by flow cytometry by means of annexin V-FITC/propidium iodide (PI) staining (BD, USA) following the manufacturer’s instructions. After treatment for 24 h, the cells were harvested and washed in cold phosphate buffer saline (PBS) twice, then double stained by Annexin V and PI in 100 μL 1 × binding buffer at room temperature for 15 min in the dark. After incubation, another 400 μL 1× annexin-binding buffer was added. The apoptotic cells were then analyzed by a flow cytometry (BD, 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!