In molecular biology, an annexin A5 affinity assay is a test to quantify the number of cells undergoing apoptosis. The assay uses the protein annexin A5 to tag apoptotic and dead cells, and the numbers are then counted using either flow cytometry or a fluorescence microscope .
Annexin V stained cells are used to indicate cell membrane changes that occur in the early stage of apoptosis. Once apoptosis is initiated, the phosphatidylserine presents in the inner cell membrane migrates through the cell membrane of the lipid bilayer. Annexin V does not attach to the cells. During early apoptosis, the PS are exposed on the outer layer, where they attach to the FITC-labeled Annexin V and stain the cell surface green. During late apoptosis, propidium iodide (PI) enters the cell and stains the contents red. Application 1 Fluorescent imaging of apoptosis induced cells Application 2 Annexin V is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner.
Annexin V: In normal viable cells, phosphatidylserine (PS) is located on the cytoplasmic surface of the cell membrane. However, in the intermediate stages of apoptosis, PS is translocated from the inner to the outer leaflet of the membrane, exposing PS to the external cellular environment where it can be detected. The kit is used for the combined detection of early-stage cell apoptosis (using Annexin V) and late-stage cell apoptosis or necrosis (using both Annexin V and 7-AAD). Annexin V belongs to the annexin family of intracellular proteins that bind to membrane phospholipids in a calcium-dependent manner.
PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during apoptosis, PS translocates to the external leaflet. The Calbiochem ® Annexin V-FITC Apoptosis Detection Kit II utilizes annexin V conjugated to FITC to bind to phosphatidylserine on the outer surface of cells undergoing apoptosis. Fluorescence is then detected by flow cytometry or fluorescence microscopy.
Annexin V binding assay as a tool to measure apoptosis in differentiated neuronal cells. We describe a rapid and reliable method to quantitate the extent of apoptosis in neuronal cell cultures.
2015-03-09 · Annexin V Conjugates for Apoptosis Detection | 3 Before Starting Fluorescence spectral characteristics The excitation and emission maxima for the various conjugates are shown in Table 1. Storage and handling The fluorescent annexin V conjugates are in a solution containing 25 mM HEPES, 140 mM NaCl, 1 mM EDTA, pH 7.4, plus 0.1% bovine serum 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. Annexin V may be conjugated to fluorochromes including FITC.
A basic protocol and the required reagents, unless otherwise noted, are found in the Annexin V Apoptosis Kit - FITC (catalog # NBP2-29373) MATERIALS > PBS (not included in the kit) > 1X Binding buffer: 10 mM Hepes pH 7.4, 140 mM NaCl and 2.5 mM CaCl 2 (10X dilution before use) > Positive control cells > Annexin V-FITC staining solution > Propidium iodine (PI) staining solution
Annexin V may also bind to PS on late apoptotic and necrotic cells but as the membrane integrity on these cells has been lost, these cells can be distinguished from early apoptotic cells by the use of the impermeant dye propidium iodide. Annexin V-FITC Apoptosis Detection Kit | Sigma-Aldrich.
Binding of Annexin V to the exposed charged head
The Annexin V corresponding signal provides a very sensitive method for detecting cellular apoptosis, while propidium iodide (PI) is used to detect necrotic or
During apoptosis, PS is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for
As such, Annexin V can be conjugated to biotin or to a fluorochrome , and used for the easy, flow cytometric identification of cells in the early stages of apoptosis. In addition, following induction of apoptosis—that is, at the early apoptotic stage —certain characteristic changes appear on the membrane. A normal cell
Annexin V is a phospholipid phosphatidylserine (PS) binding protein.
Supplier, Southern Biotechnology. Description, ApoScreen® Annexin V Apoptosis Kit-FITC.
Choose Variant. Annexin V-Biotin Detection Kit.
BioLegend's APC Annexin V Apoptosis Detection Kit with PI has been specifically designed for the identification of apoptotic and necrotic cells. Annexin V (or
29 Aug 2016 Annexin V assays allow you to detect apoptotic cells and are available in bovine, canine, equine, chicken, and swine species.
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These annexin V conjugates provide quick and reliable detection methods for studying the
Request a Quote. Annexin V-FITC staining of apoptosis in Jurkat T cells. Annexin V-FITC staining of Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that Annexin V may be conjugated to fluorochromes including Phycoerythrin (PE).
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Apoptosis and phagocytosis of tissue-dwelling eosinophils in sinonasal polyps Annexin-V-positive eosinophils were not as frequent as CD95+ cells, but
In molecular biology, an annexin A5 affinity assay is a test to quantify the number of cells undergoing apoptosis. The assay uses the protein annexin A5 to tag apoptotic and dead cells, and the numbers are then counted using either flow cytometry or a fluorescence microscope . FITC-labelled recombinant human annexin V shows bright green fluorescence (Ex(max): 488nm, Em(max): 530nm).