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 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. Bd annexin vv450 serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis manufacturer. The ability of cells to bind annexin v is still another marker considered to be specific to apoptosis. Apc annexin v apoptosis detection kit with 7aad fc ruo. Early apoptotic cells express phosphatidylserines ps on. Annexin 1, glucocorticoids, and the neuroendocrineimmune. Destroying the integrity of the plasma membrane will allow binding of annexin v to ps inside the cell.
Tacs annexin vfitc apoptosis detection kit 4830250k. The relative proportion of annexin vpositive platelets varies markedly, from a few percent 4, 9 up to 70% 10. 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. The principle of apoptosis detection method based on annexin v. However, it is like most assays and has it limitations. 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. Pi data was ok but fitc annexin v data has a problem. Anyone familiar with apoptosis interpretation n annexin vpi. Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometrybased methods. Binding of annexin v is connected with apoptotic changes in nuclearmorphology, dna fragmentation and membrane leaflet symmetry. Inappropriately regulated apoptosis is implicated in disease states, such as alzheimers disease and cancer. They are mostly found in eukaryotic organisms animal, plant and fungi.
Annexin v staining is a common method for detecting apoptotic cells. Annexin a5 or annexin v is a cellular protein in the annexin group. Annexin v staining of adherent cells for flow cytometry. Aug062009 do you know a good method to detect apoptosis on adherent cells. Apc annexin v apoptosis detection kit with pi fc ruo. One should keep in mind, however, that use of the annexin v binding assay is hindered in some instances, e. Annexin ii bd biosciences 610069 product information. All annexins contain a putative pkc binding site, but only annexin v.
Annexin v or annexin a5 is a member of the annexin family of intracellular. There are a limited number of studies investigating annexin v binding in whole. This apoptosis lecture explains about the structure and function of annexin v in apoptosis or cell death. V242 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. V351 introduction apoptosis is a carefully regulated process of cell death that occurs. Annexin v7aad staining in keratinocytes springerlink. 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.
Annexin vcy3 binding can be detected by flow cytometry ex 488 or 543 nm. I used bd fitc annexin v apoptosis detection kit 556547 and cell line is breast cancer cell. Detection of apoptosis using the bd annexin v fitc assay. 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. Annexinv and apoptosis flow cytometry core facility. Harvest cells and wash once with azide and proteinfree 1x dpbs. 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. The special engineering process ensures the brighter fluorescence signal and better photo stability. Aqueous buffered solution containing no preservative. Flow cytometry of apoptosis unit 18 university of illinois. Apc annexin v apoptosis detection kit with pi biolegend. Detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system.
Annexin v or annexin a5 is a member of the annexin family of. Using flow cytometry, titrate annexin v against the usual number of cells youll be using in your experiments, at the appropriate volume. Fitc annexin vdead cell apoptosis kit with fitc annexin v. Annexin v staining of adherent cells for flow cytometry for research use only. Annexin v can be used as a fluorescent probe to label apoptotic cells.
Fluorescent conjugates of annexin v are commonly used to identify apoptotic cells. Bd pharmingen technical data sheet pe annexin v apoptosis detection kit i product information material number. Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for. However, due to membrane disintegration during necrosis, annexin v will also bind to intracellularly located ps in necrotic cells. Annexin v binding assay as a tool to measure apoptosis in. In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of. For staining of living tissues, the specimen would need to be thin enough to allow exposure. Annexin v can only be used as a marker of apoptosis in cells where the plasma membrane is intact. Fitc annexin v apoptosis detection kit with 7aad biolegends fitc annexin v. Modified annexin vpropidium iodide apoptosis assay for. Add annexin v according to the manufacturers recommendations e.
Fitc annexin v apoptosis detection kit with 7aad biolegend. A tool for identifying cells that are undergoing apoptosis. Not for use in diagnostic or therapeutic procedures. Annexin is a common name for a group of cellular proteins. In collaboration with nexins research bv, we provide the best and brightest annexin v conjugates available, including this fitc annexin v conjugate.
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. Apc annexin vdead cell apoptosis kit with apc annexin v and. Thus, the maxpar labeling protocol would not work with annexin v, unless a free sh group can be introduced to annexin v. Thymocytes were left untrated left or treated with 100 nm dexamethasone for 15. Apoptosis can be induced by uvirradiation, as well as by numerous chemical treatments including the use of pankinase. Annexin v7aminoactinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Posttranslational modifications of annexin a2 are linked to its association with perinuclear nonpolysomal mrnp complexes. 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.
Bd pharmingen fitc annexin v apoptosis detection kit. Wash cells twice in azidefree and serumproteinfree pbs. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v. Detection of apoptotic dexamethasontreated thymocytes by annexin v staining. 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 annexin v apoptosis assay university of kansas.
A fluorescein isothiocyanate fitc conjugated form of annexin v is used to detect ps exposing cells thereby marking apoptotic cells. Annexin v binding to platelets is agonist, time and. After 6 h of induction, cells with the characteristic. The annexin v apoptosis assay development of the annexin v apoptosis assay. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to phosphatidylserine ps in a calciumdependent manner. Fitc annexin vdead cell apoptosis kit with fitc annexin v and pi, for flow cytometry catalog no.
Pdf detection of apoptosis using the bd annexin v fitc. Annexin v staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. Jurkat t cells were left untreated a or treated for 4 hr with 12 m camptothecin b. Hingorani and others published detection of apoptosis using the bd annexin v fitc assay. Pdf detection of apoptosis using the bd annexin v fitc assay on. Apoptosis was induced by culturing chp212 cells in the presence of the cyclin dependent kinase inhibitor roscovitine schutte et al. 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. Annexin v fitc staining can identify apoptosis at an earlier stage than our apobrdu or apodirect kits based on dna fragmentation in the nucleus. Microsoft powerpoint \262\323\255m\245\\\257\340\264\372\270. Dilute apoptosis reagents in desired medium formulations a. Propidium iodide pi is widely used in conjunction with annexin v to. Detection of apoptotic dexamethasonetreated thymocytes by annexin v staining. Any suggestions on the problem of too many annexin vpi. Ive read on the datasheet of bd annexin v apoptosis.
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