Product Catalog

The analysis of phosphatidylserine on the outer leaflet of apoptotic cell membranes is performed using biotin-labeled Annexin-V. Additionally, staining with propidium iodide (PI) in FACS analysis or with a cell surface marker can be used for differentiation from necrotic cells. Annexin-V-Biotin can be used in immunohistochemistry with a secondary detection system (e.g., Streptavidin-POD conjugate).

Detection: Annexin-V-Biotin can be used in immunohistochemistry with a secondary detection system (e.g., Streptavidin-POD conjugate).
Sample material: Cell lines and freshly isolated cells
Specificity: Annexin-V binds in a calcium-dependent manner to negatively charged phospholipid surfaces, and shows high affinity for phosphatidylserine.

In the early stages of apoptosis there are changes occurring on the cell surface, specifically translocation of phosphatidylserine (PS) from the interior side of the plasma membrane to the outer leaflet. This secretion of phosphatidylserine is the ideal basis for Annexin-V, which binds to PS with high affinity. Annexin-V is a Ca²⁺-dependent phospholipid-binding protein with a high affinity for PS. Since necrotic cells also expose PS as a result of lost membrane integrity, apoptotic cells must be differentiated from these necrotic cells. The simultaneous application of a DNA stain, which is used for dye-exclusion testing, allows the discrimination of necrotic cells from the Annexin-V positively stained cell cluster.

Contents
500 µl, ready-to-use solution, for 250 tests

1. Induce apoptosis.
2. Wash the cells in PBS.
3. Incubate cells with Annexin-V-Biotin in a Hepes buffer that contains PI or other labeling reagent for cell surfaces.
4. Analyze samples under a microscope or on a flow cytometer.