I would like to analyse apoptosis by flow cytometry.
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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. The annexin a5 protein binds to apoptotic cells in a calcium-dependent manner using phosphatidylserine-containing.
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Annexin A5 binds to biological membranes, exposing negatively charged phospholipid in a calcium-dependent manner, self-assembling into ordered two-dimensional arrays on membrane surfaces. 71 Antiphospholipid antibodies have been shown to disrupt this annexin A5 lattice, or “shield,” exposing the anionic phospholipid membranes beneath, which can support phospholipid-dependent coagulation.
Analysis of the membrane-bound form of annexin V by electron microscopy revealed no differences between the wild type and mutant. Electrophysiology and structural studies were performed on an annexin V variant containing a mutation of glutamic acid-95 to serine in the center of the pore region. The mutation resulted in a lower single channel conductance for calcium and a strongly increased.
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