Staurosporine was purchased from Sigma-Aldrich (Roche Diagnostics GmbH), and the Annexin V kit from BioVision (Milpitas, CA). W805EC NE, an oil-in-water emulsion made from ingredients that are Generally Recognized As Safe (GRAS) was provided by the NanoBio Corporation (Ann Arbor, MI). Caspases 1, 6, 8, 9 and 3/7 were assessed using caspase kits (ImmunoChemistry Technologies, Bloomington, MN) and the CaspaTag Caspase 3/7 kit (EMD Millipore, MA), respectively.
Annexin 5 kit
Manufactured by Abcam
Sourced in United States
The Annexin V kit is a laboratory reagent used for the detection and quantification of apoptosis. It functions by binding to phosphatidylserine, a molecule that is externalized on the surface of cells undergoing programmed cell death.
Automatically generated - may contain errors
5 protocols using annexin 5 kit
1
Apoptosis Induction and Caspase Activation
2
Phosphatidylserine Externalization and Caspase Activity
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Analyzing Cell Cycle and Apoptosis in NSCLC
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Human NSCLC cells were seeded onto 100 mm dishes with DMEM medium and grown for 24 h for stabilization. Then, cells were treated and incubated with PB01 (100 nM) or DMSO for 24 h. For cell cycle analysis, NSCLC cells were fixed with cold 80% ethanol and stained with a solution containing PI and RNase A (Sigma). The ROS generation during apoptosis was examined by monitoring the cells after staining with the cell-permeant 2′7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA, Invitrogen). For annexin V analysis, cells were fixed and stained annexin V/PI dye for 30 min using annexin V kit (Biovision, Palo Alto, CA, USA). Data were analyzed with a FACS Calibur flow cytometer (Becton Dickinson, San Jose, CA, USA).
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Annexin V Flow Cytometry Apoptosis Assay
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An annexin V kit (BioVision Inc. USA) was used to determine the percentage of cells undergoing apoptosis. Briefly, the CMECs were harvested after 24-hour treatments. Approximately, 15.000 cells were detected for each sample. Then, the cells were trypsinized gently and resuspended with binding buffer, and they were treated with 5 µL annexin V–Fluorescein Isothiocyanate (FITC) and 5 µL propidium iodide (PI). After the incubation for 5 min on ice, each sample was analyzed immediately using the FACSCalibur flow cytometer (Becton Dickinson, USA).
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Annexin-V Assay for Protozoan Apoptosis
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Here, to study the effects of SLME on the detection of apoptotic and necrotic cells in T. gondii tachyzoites, we used the Annexin-V Kit (BioVision, Waltham, MA, USA) based on the producer’s instructions. In summary, tachyzoites (1 × 106 cells/mL) were incubated with SLME at 32.5, 75, and 150 µg/mL in 24-well plates for 48 h at 24 °C. Followed by centrifuging at 2500 rpm and discarding the upper phase, we added the kit materials, including binding buffer, Annexin-V58T, and propidium iodide (PI) to the wells, and were incubated in dark condition for 5 min at 21 °C. Finally, absorbance in cells was measured at the excitation wavelength of 570 nm and the emission wavelength of 630 nm by flow cytometry (Sysmex Partec GmbH, Munster, Germany), and the obtained findings were examined by FlowJo software.
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