What does annexin V measure?
What does annexin V measure?
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 is annexin V stain?
Annexin V staining is a common method for detecting apoptotic cells. Thermo Fisher Scientific offers high-quality fluorescent annexin V conjugates as standalone reagents and in a variety of kits for use in flow cytometry and for imaging suspension cells.
Can annexin V be fixed?
Yes, cells can be fixed with formaldehyde after staining. Because Annexin V staining is dependent on calcium, all buffers used for washing and fixation should contain 1.25 mM CaCl2. Fixation may increase the background signal from Ethidium Homodimer III.
What is Annexin PI assay?
The Annexin V/ PI protocol is a commonly used approach for studying apoptotic cells3. In late apoptotic and necrotic cells, the integrity of the plasma and nuclear membranes decreases7,8, allowing PI to pass through the membranes, intercalate into nucleic acids, and display red fluorescence 1,2,9.
What is Annexin V FITC?
General description. Annexin V-FITC kit allows fluorescent detection of annexin V bound to apoptotic cells and quantitative determination by flow cytometry. The AnnexinV-FITC kit uses annexin V conjugated with fluorescein isothiocyante (FITC) to label phosphatidylserine sites on the membrane surface.
What is in binding buffer?
The binding buffer is an isotonic buffer containing calcium, which is essential for the binding of Annexin V to phosphatidylserine.
How does annexin V-FITC work?
Annexin V is a 35-36 kDa Ca2+ dependent phospholipid-binding protein that has a high affinity for PS, and binds to cells with exposed PS. While cells that are in early apoptosis are Annexin V-FITC positive and PI negative and cells that are in late apoptosis or already dead are both FITC Annexin V and PI positive.
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).
What is FITC used for?
Fluorescein isothiocyanate (FITC) is widely used to attach a fluorescent label to proteins vi a the amine group. The isothiocyanate group reacts with amino terminal and primary amines in proteins. It has been used for the labeling of proteins including antibodies and lectins.
