In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of. Hingorani and others published detection of apoptosis using the bd annexin v fitc assay. Propidium iodide pi is widely used in conjunction with annexin v to. Detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system bd biosciences august 2011 introduction apoptosis is a normal genetically programmed process that occurs during embryonic development, as well as in maintenance of tissue homeostasis, under pathological conditions, and in aging. Aqueous buffered solution containing no preservative. The principle of apoptosis detection method based on annexin v. Annexin 1, glucocorticoids, and the neuroendocrineimmune. Annexin v can only be used as a marker of apoptosis in cells where the plasma membrane is intact. Apc annexin v apoptosis detection kit with 7aad biolegends apc annexin v apoptosis detection kit with 7aad has been specifically designed for the identification of apoptotic and necrotic cells. Thymocytes were left untrated left or treated with 100 nm dexamethasone for 15. Apoptosis was induced by culturing chp212 cells in the presence of the cyclin dependent kinase inhibitor roscovitine schutte et al. Inappropriately regulated apoptosis is implicated in disease states, such as alzheimers disease and cancer.
Annexin v, also known as pap1 or lipocortin v, is a 30 kda protein that binds to phospholipids in a calciumdependent manner 1. Detection of apoptotic dexamethasonetreated thymocytes by annexin v staining. Cells were incubated with annexin v fitc in a buffer containing propidium iodide pi, cat. Bd annexin vv450 serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis manufacturer. Fitc annexin vdead cell apoptosis kit with fitc annexin v. Fitc annexin v is a sensitive probe for identifying apoptotic cells, binding to negatively charged phospholipid surfaces kd of 5 x 10e2 with a higher affinity for phosphatidylserine ps than most other phospholipids. Detection of apoptosis using annexin v and the nc3000.
Annexin v7aad staining in keratinocytes springerlink. Annexin v can only be used as a marker of apoptosis in cells where the plasma membrane is intact because destroying the integrity of the plasma membrane will allow nonspecific binding of annexin v. Measuring apoptosis using annexin v and flow cytometry. Annexin v staining of adherent cells for flow cytometry. Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for.
Tacs annexin vfitc apoptosis detection kit 4830250k. The annexin v apoptosis assay development of the annexin v apoptosis assay. Wash cells twice in azidefree and serumproteinfree pbs. This apoptosis lecture explains about the structure and function of annexin v in apoptosis or cell death. All annexins contain a putative pkc binding site, but only annexin v. Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for imaging and flow cytometry instrument platforms. Pi data was ok but fitc annexin v data has a problem.
In collaboration with nexins research bv, we provide the best and brightest annexin v conjugates available, including this fitc annexin v conjugate. Detection of apoptosis using the bd annexin v fitc assay. Detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system. Annexin v7aminoactinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Annexin v binding to platelets is agonist, time and. Annexin v staining of adherent cells for flow cytometry for research use only. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to phosphatidylserine ps in a calciumdependent manner. Annexin v, belonging to a recently discovered family of proteins, the annexins, with anticoagulant properties has proven to be a useful tool in detecting apoptotic cells since it preferentially binds to.
Annexin v binding assay as a tool to measure apoptosis in. Destroying the integrity of the plasma membrane will allow binding of annexin v to ps inside the cell. Annexinv and apoptosis flow cytometry core facility. Annexin v staining is a common method for detecting apoptotic cells.
Annexin v can be used as a fluorescent probe to label apoptotic cells. Anyone familiar with apoptosis interpretation n annexin vpi. V351 introduction apoptosis is a carefully regulated process of cell death that occurs. They are mostly found in eukaryotic organisms animal, plant and fungi. Annexin v staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. Posttranslational modifications of annexin a2 are linked to its association with perinuclear nonpolysomal mrnp complexes. Pdf detection of apoptosis using the bd annexin v fitc assay on. For staining of living tissues, the specimen would need to be thin enough to allow exposure. Annexin ii bd biosciences 610069 product information. Apc annexin v apoptosis detection kit with pi biolegend. Hingorani and others published detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system find, read and cite all the research you need on.
Harvest cells and wash once with azide and proteinfree 1x dpbs. Thus, the maxpar labeling protocol would not work with annexin v, unless a free sh group can be introduced to annexin v. Apc annexin v apoptosis detection kit with pi fc ruo. Annexin v or annexin a5 is a member of the annexin family of.
However, it is like most assays and has it limitations. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v. Early apoptotic cells express phosphatidylserines ps on. The annexin v apoptosis assay university of kansas. Apc annexin vdead cell apoptosis kit with apc annexin v and.
Apoptosis can be induced by uvirradiation, as well as by numerous chemical treatments including the use of pankinase. After 6 h of induction, cells with the characteristic. One should keep in mind, however, that use of the annexin v binding assay is hindered in some instances, e. Fitc annexin v apoptosis detection kit with 7aad biolegend. A tool for identifying cells that are undergoing apoptosis. Annexinv is a specific psbinding protein that can be used to detect apoptotic cells. Not for use in diagnostic or therapeutic procedures. However, due to membrane disintegration during necrosis, annexin v will also bind to intracellularly located ps in necrotic cells. V242 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Apc annexin v apoptosis detection kit with 7aad fc ruo. Fitc annexin v apoptosis detection kit with 7aad biolegends fitc annexin v. Binding of annexin v is connected with apoptotic changes in nuclearmorphology, dna fragmentation and membrane leaflet symmetry. There are a limited number of studies investigating annexin v binding in whole.
Annexin v apoptosis detection assay kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidylserine ps from the inner face of the plasma membrane. Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometrybased methods. Annexin v or annexin a5 is a member of the annexin family of intracellular. Fitc annexin vdead cell apoptosis kit with fitc annexin v and pi, for flow cytometry catalog no. Ive read on the datasheet of bd annexin v apoptosis. Using flow cytometry, titrate annexin v against the usual number of cells youll be using in your experiments, at the appropriate volume. Any suggestions on the problem of too many annexin vpi. Pdf detection of apoptosis using the bd annexin v fitc. Bd pharmingen fitc annexin v apoptosis detection kit. Annexin vcy3 binding can be detected by flow cytometry ex 488 or 543 nm. Dilute apoptosis reagents in desired medium formulations a. Annexin v fitc staining can identify apoptosis at an earlier stage than our apobrdu or apodirect kits based on dna fragmentation in the nucleus. I used bd fitc annexin v apoptosis detection kit 556547 and cell line is breast cancer cell. A fluorescein isothiocyanate fitc conjugated form of annexin v is used to detect ps exposing cells thereby marking apoptotic cells.
Fluorescent conjugates of annexin v are commonly used to identify apoptotic cells. Fitc apoptosis detection kit i staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from either apoptotic or necrotic processes. The ability of cells to bind annexin v is still another marker considered to be specific to apoptosis. Flow cytometry of apoptosis unit 18 university of illinois. Bd pharmingen technical data sheet pe annexin v apoptosis detection kit i product information material number. The relative proportion of annexin vpositive platelets varies markedly, from a few percent 4, 9 up to 70% 10. Jurkat t cells were left untreated a or treated for 4 hr with 12 m camptothecin b. Detection of apoptotic dexamethasontreated thymocytes by annexin v staining. Annexin is a common name for a group of cellular proteins. Add annexin v according to the manufacturers recommendations e. Modified annexin vpropidium iodide apoptosis assay for. Microsoft powerpoint \262\323\255m\245\\\257\340\264\372\270. The special engineering process ensures the brighter fluorescence signal and better photo stability.
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