Aqueous buffered solution containing no preservative. Annexin v can be used as a fluorescent probe to label apoptotic cells. Tacs annexin vfitc apoptosis detection kit 4830250k. Bd pharmingen technical data sheet pe annexin v apoptosis detection kit i product information material number. In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of. In collaboration with nexins research bv, we provide the best and brightest annexin v conjugates available, including this fitc annexin v conjugate.
Fitc annexin vdead cell apoptosis kit with fitc annexin v. Apc annexin v apoptosis detection kit with pi biolegend. Annexin vcy3 binding can be detected by flow cytometry ex 488 or 543 nm. The special engineering process ensures the brighter fluorescence signal and better photo stability. Annexin v7aminoactinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Annexin v binding assay as a tool to measure apoptosis in. Posttranslational modifications of annexin a2 are linked to its association with perinuclear nonpolysomal mrnp complexes. After 6 h of induction, cells with the characteristic. Annexinv and apoptosis flow cytometry core facility. Ive read on the datasheet of bd annexin v apoptosis.
Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for. Detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system. They are mostly found in eukaryotic organisms animal, plant and fungi. I used bd fitc annexin v apoptosis detection kit 556547 and cell line is breast cancer cell. Annexin v staining is a common method for detecting 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. Pdf detection of apoptosis using the bd annexin v fitc. 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.
All annexins contain a putative pkc binding site, but only annexin v. V351 introduction apoptosis is a carefully regulated process of cell death that occurs. Pi data was ok but fitc annexin v data has a problem. There are a limited number of studies investigating annexin v binding in whole. Annexin 1, glucocorticoids, and the neuroendocrineimmune. Apoptosis can be induced by uvirradiation, as well as by numerous chemical treatments including the use of pankinase. Annexin is a common name for a group of cellular proteins. 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 pi fc ruo. Annexin v staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. Bd annexin vv450 serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis manufacturer. Add annexin v according to the manufacturers recommendations e.
Destroying the integrity of the plasma membrane will allow binding of annexin v to ps inside the cell. Apc annexin v apoptosis detection kit with 7aad fc ruo. Annexin v or annexin a5 is a member of the annexin family of. Annexin a5 or annexin v is a cellular protein in the annexin group. Fluorescent conjugates of annexin v are commonly used to identify apoptotic cells. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to phosphatidylserine ps in a calciumdependent manner.
Anyone familiar with apoptosis interpretation n annexin vpi. Early apoptotic cells express phosphatidylserines ps on. The ability of cells to bind annexin v is still another marker considered to be specific to apoptosis. The relative proportion of annexin vpositive platelets varies markedly, from a few percent 4, 9 up to 70% 10. Annexin ii bd biosciences 610069 product information. Annexin v staining of adherent cells for flow cytometry. Annexinv is a specific psbinding protein that can be used to detect apoptotic cells. Any suggestions on the problem of too many annexin vpi.
Thus, the maxpar labeling protocol would not work with annexin v, unless a free sh group can be introduced to annexin v. 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. The principle of apoptosis detection method based on annexin v. A tool for identifying cells that are undergoing apoptosis. 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. However, it is like most assays and has it limitations. Detection of apoptotic dexamethasontreated thymocytes by annexin v staining. Jurkat t cells were left untreated a or treated for 4 hr with 12 m camptothecin b.
V242 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Microsoft powerpoint \262\323\255m\245\\\257\340\264\372\270. Flow cytometry of apoptosis unit 18 university of illinois. Annexin v7aad staining in keratinocytes springerlink. A fluorescein isothiocyanate fitc conjugated form of annexin v is used to detect ps exposing cells thereby marking apoptotic cells. 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. Using flow cytometry, titrate annexin v against the usual number of cells youll be using in your experiments, at the appropriate volume. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v. One should keep in mind, however, that use of the annexin v binding assay is hindered in some instances, e.
Fitc annexin vdead cell apoptosis kit with fitc annexin v and pi, for flow cytometry catalog no. Modified annexin vpropidium iodide apoptosis assay for. However, due to membrane disintegration during necrosis, annexin v will also bind to intracellularly located ps in necrotic cells. Annexin v or annexin a5 is a member of the annexin family of intracellular. Thymocytes were left untrated left or treated with 100 nm dexamethasone for 15. This apoptosis lecture explains about the structure and function of annexin v in apoptosis or cell death. 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. Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometrybased methods. Inappropriately regulated apoptosis is implicated in disease states, such as alzheimers disease and cancer. Wash cells twice in azidefree and serumproteinfree pbs.
For staining of living tissues, the specimen would need to be thin enough to allow exposure. Annexin v fitc staining can identify apoptosis at an earlier stage than our apobrdu or apodirect kits based on dna fragmentation in the nucleus. Annexin v staining of adherent cells for flow cytometry for research use only. The annexin v apoptosis assay university of kansas.
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. Dilute apoptosis reagents in desired medium formulations a. 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. Detection of apoptosis using annexin v and the nc3000. 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.
Aug062009 do you know a good method to detect apoptosis on adherent cells. Binding of annexin v is connected with apoptotic changes in nuclearmorphology, dna fragmentation and membrane leaflet symmetry. Propidium iodide pi is widely used in conjunction with annexin v to. 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 apoptotic dexamethasonetreated thymocytes by annexin v staining. Detection of apoptosis using the bd annexin v fitc assay. Apc annexin vdead cell apoptosis kit with apc annexin v and. Apoptosis was induced by culturing chp212 cells in the presence of the cyclin dependent kinase inhibitor roscovitine schutte et al. Annexin v, also known as pap1 or lipocortin v, is a 30 kda protein that binds to phospholipids in a calciumdependent manner 1.
Bd pharmingen fitc annexin v apoptosis detection kit. Fitc annexin v apoptosis detection kit with 7aad biolegends fitc annexin v. Cells were incubated with annexin v fitc in a buffer containing propidium iodide pi, cat. 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.
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