CLL lymphocyte cells were left untreated or treated with DMSO alone; 0.5 nM ABT-199 or 1 nM ABT-737; 3 or 10 μM AZD1208, SMI-4a, or SGI-1776; or a combination of a Bcl-2 antagonist and a Pim kinase inhibitor at the indicated concentrations. Cells were washed, resuspended in 200 μL of Annexin binding buffer (Roche, Indianapolis, IN), mixed with 5 μL of Annexin V solution (BD Pharmingen, San Diego, CA) plus 5 μL of propidium iodide (PI; Sigma-Aldrich, St. Louis, MO), and incubated for 15 min in the dark at room temperature. At least 1 x 104 cells were measured per sample using a Becton Dickinson FACSCalibur flow cytometer (San Jose, CA).
Annexin binding buffer
Manufactured by Roche
Sourced in United States
Annexin binding buffer is a solution used in flow cytometry experiments to measure apoptosis. It provides the necessary ionic conditions for the binding of annexin V to phosphatidylserine, which is exposed on the surface of apoptotic cells.
Automatically generated - may contain errors
2 protocols using annexin binding buffer
1
Annexin V and PI Apoptosis Assay
2
Annexin V/PI Apoptosis Assay
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Cells were untreated or treated with DMSO or AZD1208 and cell death (Annexin V/propidium iodide (PI) positivity) was measured using a Becton Dickinson FACSCalibur flow cytometer (Franklin Lakes, NJ, USA). Briefly, cells were washed, suspended in Annexin binding buffer (200 μL) (Roche; Indianapolis, IN, USA), mixed with Annexin V solution (5 μL) (BD Pharmingen; San Diego, CA, USA) plus PI (5 μL) (Sigma-Aldrich; St. Louis, MO, USA) and incubated for 15 min in the dark at room temperature. At least 10,000 cells were measured per sample.
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