Cell culture

Apoptosis

Annexin V-Fluorescein – 494nm/521nm PI – 536nm/617nm

Flow cytometry

Porcine Annexin V-Fluorescein Apoptosis BioAssay™ Kit provides a proven method for quickly and easily distinguishing two populations of dying cells from viable cells using recombinant fluorescein-conjugated Swine Annexin V and Propidium Iodide (PI). These cells will become dually labeled with Annexin V-Fluorescein (green fluorescence) and Propidium Iodide (red fluorescence). Results can be analyzed via flow cytometry.

Annexin V is a member of a calcium and phospholipid binding family of proteins with vascular anticoagulant activity. Results from in vitro experiments indicate that it may play a role in the inhibition of blood coagulation by competing for phosphatidylserine (PS) binding sites with prothrombin. In healthy cells, PS is usually kept in the inner-leaflet (the cytosolic side) of the cell membrane. When a cell undergoes apoptosis, one of the earliest detectable indicators is the loss of membrane asymmetry. No longer restricted to the cytosolic part of the membrane, PS is translocated to the outer-leaflet and becomes exposed on the surface of the cell.

Porcine Annexin V-Fluorescein Apoptosis BioAssay™ Kit provides a proven method for quickly and easily distinguishing two populations of dying cells from viable cells using recombinant fluorescein-conjugated Porcine Annexin V and Propidium Iodide (PI). Cells with intact cell membranes and surface-exposed PS, a prominent feature of apoptosis, will stain positive for Annexin V-Fluorescein. PI is included in the BioAssay™ kit to identify dying, later stage apoptotic cells which have lost plasma membrane integrity. These cells will become dually labeled with Annexin V-Fluorescein and Propidium Iodide (green and red fluorescence). Live non-apoptotic cells with intact plasma membranes will exclude PI and will remain unstained by the Annexin V-Fluorescein probe, assuming no treatment or cell cycle-associated event temporarily exposes the normally internalized, negatively charged PS entity. The BioAssay™ kit also includes a specially formulated, calcium-based binding buffer which is required for Annexin V binding to occur. Analyze results using a flow cytometry.

Porcine Annexin V-Fluorescein, lyophilized, 1 vial Annexin Reconstitution Buffer, 1X, 1x1mL, 1 vial Annexin Binding Buffer, 10X, 1x10mL, 3 bottles Propidium Iodide, 1x250ug/mL, 1x1mL, 1 vial

Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.