U937 cells (2.5×105 cells/ml) were incubated with piperlongumine (0–20 µM) for 48 h at 37°C in the presence of 5% CO2. Cells were washed twice with PBS and resuspended in 100 µl of Annexin V binding buffer (pH 7.4) (BD Biosciences, Franklin Lakes, NJ, USA). Then, annexin V-fluorescein isothiocyanate (BD Biosciences) was added and incubated for 15 min under dark conditions. Propidium iodide (0.1 µg/ml; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was added just prior to signal acquisition. The apoptosis rate was acquired using a FACSAria flow cytometer (BD Biosciences) and analyzed with FACSDiva 7.6.1 software (BD Biosciences).
Annexin 5 binding buffer ph 7.4
Manufactured by BD
Sourced in United States
Annexin V binding buffer (pH 7.4) is a laboratory reagent designed to facilitate the binding of Annexin V to phosphatidylserine residues exposed on the surface of cells undergoing apoptosis. It maintains a neutral pH of 7.4 to provide optimal conditions for this binding process.
Automatically generated - may contain errors
2 protocols using annexin 5 binding buffer ph 7.4
1
Apoptosis Assay of U937 Cells
2
Apoptosis Analysis of Cisplatin and ABT-737 Treated Cells
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The M2 cells were seeded on 6-well plates at a density of 1.25 × 105 cells per well. The following day they were exposed to 40 μM cisplatin or 10 μM ABT-737 for 24 h. A375 cells were treated with 10 μM ABT-737 and 40 μM ML258, either alone or in combination for 24 h. Cells were washed twice with PBS and resuspended in 100 µL of annexin V binding buffer (pH 7.4) (BD Biosciences, Franklin Lakes, NJ, USA). Then, annexin V-Alexa Fluor 488 (BD Biosciences) was added and incubated for 15 min under dark conditions. Propidium iodide (0.1 µg/mL; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was added just prior to signal acquisition. Cells were analyzed using a FACSAria flow cytometer (BD Biosciences, San Jose, CA) and analyzed with FACSDiva 7.6.1 software (BD Biosciences).
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