FACS was also used to determine the proportion of cells entering apoptosis following chrysin treatment using a PE Annexin V and 7-AAD Apoptosis Detection Kit I (BD Pharmingen, San Jose, CA, USA). Cells were harvested using Cellstripper (Cellgro, Manassas, VA, USA), a nonenzymatic cell dissociation solution, and floating cells were harvested as well. Cells were pelleted and resuspended in 1× PBS twice and then resuspended in binding buffer (10mM HEPES/NaOH, pH=7.4, 140mM NaCl, and 2.5mM CaCl2) at 1×106 cells/ml. Next, cell suspensions were incubated in PE Annexin V and 7-AAD (7 Aminoactinomycin D) fluorescein solutions for 15 minutes in the dark at room temperature and sorted using the FACSCalibur™. Cell distributions were determined using FlowJo v10.0.8 software (TreeStar, Inc., Ashland, OR, USA). Three biological replicates of this experiment are presented as a mean ± SEM.
Pe annexin 5 and 7 aad apoptosis detection kit 1
Manufactured by BD
Sourced in United States
The BD PE Annexin V and 7-AAD Apoptosis Detection Kit I is a laboratory reagent used to detect and analyze apoptosis, a type of programmed cell death. The kit contains Annexin V conjugated with the fluorescent dye phycoerythrin (PE) and the DNA-binding dye 7-aminoactinomycin D (7-AAD).
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2 protocols using pe annexin 5 and 7 aad apoptosis detection kit 1
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Apoptosis Quantification by FACS Analysis
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Apoptosis Quantification by FACS Analysis
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FACS was also used to determine the proportion of cells entering apoptosis following chrysin treatment using a PE Annexin V and 7-AAD Apoptosis Detection Kit I (BD Pharmingen, San Jose, CA, USA). Cells were harvested using Cellstripper (Cellgro, Manassas, VA, USA), a nonenzymatic cell dissociation solution, and floating cells were harvested as well. Cells were pelleted and resuspended in 1× PBS twice and then resuspended in binding buffer (10mM HEPES/NaOH, pH=7.4, 140mM NaCl, and 2.5mM CaCl2) at 1×106 cells/ml. Next, cell suspensions were incubated in PE Annexin V and 7-AAD (7 Aminoactinomycin D) fluorescein solutions for 15 minutes in the dark at room temperature and sorted using the FACSCalibur™. Cell distributions were determined using FlowJo v10.0.8 software (TreeStar, Inc., Ashland, OR, USA). Three biological replicates of this experiment are presented as a mean ± SEM.
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