What does annexin V do?

Fluorescent conjugates of annexin V are commonly used to identify apoptotic cells. The human vascular anticoagulant annexin V is a 35–36 kDa, Ca2+-dependent phospholipid-binding protein that has a high affinity for the anionic phospholipid phosphatidylserine (PS).

Is annexin V specific for apoptosis?

Staining with annexin V, which detects the externalization of phosphatidylserine on cells, is a widespread method for the assessment of cell death, and in particular, apoptosis.

How does annexin V assay work?

Assay Principle: Annexin V binds specifically to phosphatidylserine and labelled Annexin V can be used detect apoptotic cells. Binding of Annexin V to the exposed charged head groups of PS is a Ca2+ dependent process. Propidium Iodide is used in conjunction with labelled Annexin V.

Can annexin V be fixed?

In my experience, it is possible to fix annexin V stained cells and see great results on the flow. Stain cells with annexin V (in calcium-rich binding buffer). Wash cells in binding buffer to remove unbound AxV. Fix in your choice of fixative (diluted in binding buffer).

Where is annexin V located?

Annexin A5 (or annexin V) is a cellular protein in the annexin group. In flow cytometry, annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.

What color is annexin V?

Cells that have bound annexin V-FITC will show green staining in the plasma membrane. Cells that have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane).

How do you calculate apoptosis?

Apoptosis is detected by measuring the externalization of phosphatidylserine on the plasma membrane using fluorescent-tagged annexin V. Additionally, flow cytometry can be employed to determine alterations in cell size (Bortner and Cidlowski, 2001; Warnes et al.

What Colour is annexin V?

What kind of staining does annexin V show?

Cells that have bound annexin V-FITC will show green staining in the plasma membrane. Cells that have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane). ​​. The above is an example protocol for PS exposure detection using annexin V,

How is annexin V used to detect apoptosis?

Annexin V has a strong, Ca2+-dependent affinity for PS and therefore can be used as a probe for detecting apoptosis. For robust results, we recommend our range of one-step Annexin V staining/detection kits and reagents for use in flow cytometry and cell imaging.

Is the annexin V binding assay applicable to differentiated neuronal cells?

In the present paper, we show that the annexin V-binding assay is also applicable to differentiated neuronal cells with fragile neurite outgrowths. MeSH terms

What is the binding buffer for annexin V?

Annexin V is commercially available, conjugated to most common fluorochromes. Propidium iodide (PI) Binding buffer: 10 mM Hepes adjusted to pH 7.4, 140 mM NaCl and 2.5 mM CaCl2 Always titer the Annexin V and PI to determine the correct concentrations for your assays. Induce apoptosis in cells using the desired method.