After initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate (Cy3) of Annexin V, a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. The Annexin V-Cy3 Apoptosis Detection Kit provides annexin V-Cy3, SYTOX green dye, and binding buffer. The SYTOX green dye is impermeant to live cells and apoptotic cells, but stains necrotic cells with intense green fluorescence by binding to cellular nucleic acids. After staining a cell population with annexin V-Cy3 and SYTOX Green dye in the provided binding buffer, apoptotic cells show red fluorescence, dead cells show green fluorescence and live cells show little or no fluorescence. These populations can easily be distinguished by Fluorescence microscopy using FITC and rhodamine filters or by flow cytometry using the FL1 channel (Ex=488 nm/Em=530 nm) for SYTOX Green dye and FL2 channel for Annexin V-Cy3 (Ex=543 nm/Em=570 nm).
Annexin V Cy3 Conjugate, 500µl
SYTOX Green Dye, 100 µl
10X Binding Buffer, 20 ml
Flow Cytometry, Fluorescence Microscopy
20 tests ; 50 tests ; 100 tests
Store between 2°C-8°C. Do not freeze.
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