For Annexin V apoptosis assay (BD, Pharmingen, EremBodegem, Belgium), we stained cells simultaneously with fluorescein isothiocyanate (FITC)-Annexin V and non-vital dye PI according to the manufacturer's instructions. The percentage of the M-phase cells was determined by staining with PI and antibody to phospho-histone H3 (P-H3) (Cell Signaling, Beverly, MA, USA), followed by FITC-conjugated secondary antibody (Jackson Immunoresearch Laboratories, West Grove, PA, USA). Samples were analyzed with a CyAnTM ADP flow cytometer (Dako Cytomation, Glostrup, Denmark) using an argon-ion laser tuned to 488 nm measuring forward and orthogonal light scatter. Data were acquired by Summits software and analyzed with Modfits software.
Annexin 5 apoptosis assay
Manufactured by BD
Sourced in Belgium
The Annexin V apoptosis assay is a laboratory technique used to detect and quantify apoptosis, a programmed cell death process. The assay utilizes Annexin V, a calcium-dependent phospholipid-binding protein, to identify cells undergoing apoptosis by binding to phosphatidylserine exposed on the outer cell membrane.
Automatically generated - may contain errors
4 protocols using annexin 5 apoptosis assay
1
Annexin V Apoptosis Assay Protocol
2
Cytotoxicity of Functionalized Liposomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colonic carcinoma (COLO205) and aggressive breast carcinoma (MDA-MB-231) cell lines were obtained from ATCC and used to determine the cytotoxicity of our therapeutic formulation in benchtop experiments. The cells were cultured in 12-multiwell plates at a density of 200 000 cells per mL for 24 h. The culture medium was replaced with serum-free media and 20 μL of functionalized liposomes were added. We changed to serum-free media to maintain the number of cells equal between samples. The viability of cells was measured using an Annexin V apoptosis assay (BD) following 24 h of incubation. In this assay, the cells were removed from the culture plates and stained following the manufacturer's protocol. Following this, the viable cell percentage was determined using flow cytometry.
3
Cell Cycle and Apoptosis Analysis
4
Cell Cycle and Apoptosis Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cell cycle analysis, cells were harvested, washed with PBS and fixed with 80%, −20°C ethanol for 30 minutes on ice. Cells were then washed with PBS and stained with propidium iodide/RNase solution (BD Biosciences) for 15-30 minutes at 37°C in subdued light. Cells were subsequently filtered to ensure a single cell population and analyzed using a BD Fortessa cytometer with FACS Diva software (BD Biosciences).
Annexin V apoptosis assays were performed and analyzed in accordance with the manufacturer’s instructions (BD Bioscience). For transferable resistance assays, parental cells expressing the GFP protein in the pLEX-307 backbone were mixed 1:1 with non-fluorescent cells. Cells were sorted based on GFP status for subsequent cell cycle analysis or western blotting.
Annexin V apoptosis assays were performed and analyzed in accordance with the manufacturer’s instructions (BD Bioscience). For transferable resistance assays, parental cells expressing the GFP protein in the pLEX-307 backbone were mixed 1:1 with non-fluorescent cells. Cells were sorted based on GFP status for subsequent cell cycle analysis or western blotting.
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!