What is annexin V binding?

Annexin V binds is a phospholipid phosphatidylserine (PS) binding protein. In the earlier stages of apoptosis, PS is translocated to the outer sections of the cell plasma membrane and is exposed to the external cellular environment. The mark of the late stages of cellular death is the loss of membrane integrity.

How does annexin V staining work?

Annexin V, a 36-kDa calcium-binding protein, binds to PS; therefore, fluorescently labeled Annexin V can be used to detect PS that is exposed on the outside of apoptotic cells. Annexin V can also stain necrotic cells because these cells have ruptured membranes that permit Annexin V to access the entire plasma membrane.

What is Annexin used for?

The Annexin V/ PI protocol is a commonly used approach for studying apoptotic cells3. PI is used more often than other nuclear stains because it is economical, stable and a good indicator of cell viability, based on its capacity to exclude dye in living cells 4,5.

How do you use annexin V?

Add 5 μL of fluorochrome-conjugated Annexin V to 100 μL of the cell suspension. Incubate 10-15 minutes at room temperature. Protect from light. Add 2 mL 1X binding buffer and centrifuge at 400-600 x g for 5 minutes at room temperature.

Why do I need to use annexin V binding buffer?

Why do I need to use Annexin V Binding Buffer? Annexin V binding requires the presence of calcium in the solution. So, we provide Annexin V Binding Buffer (cat # 422201), which is optimized for the best performance of Annexin V staining.

What is the recommended concentration of annannexin V binding buffer?

Annexin V Binding Buffer is recommended for use with Annexin V reagents. Cells should be resuspended in Annexin V Binding Buffer at a concentration 1×106/ml and then combined with Annexin V reagents at the recommended volumes. Application Notes

Are there any citations for the ebioscience™ annexin V apoptosis detection kit?

Not for use in diagnostic procedures. There are no frequently asked questions yet. There are no citations & references. eBioscience™ Annexin V Apoptosis Detection Kit eBioscience™ Annexin V-Biotin Apoptosis Detecti…

Can fixable viability dye eFluor 450 be used with annexin V apoptosis detection kits?

Note: Fixable Viability Dye eFluor 450 is not recommended for use with Annexin V Apoptosis Detection Kits. Annexins are a family of calcium-dependent phospholipid-binding proteins that preferentially bind phosphatidylserine (PS). Under normal physiologic conditions, PS is predominantly located in the inner leaflet of the plasma membrane.