Apc annexin v apoptosis detection kit with pi fc ruo. V242 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. Posttranslational modifications of annexin a2 are linked to its association with perinuclear nonpolysomal mrnp complexes. After 6 h of induction, cells with the characteristic.
However, it is like most assays and has it limitations. For staining of living tissues, the specimen would need to be thin enough to allow exposure. Pdf detection of apoptosis using the bd annexin v fitc. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v. Dilute apoptosis reagents in desired medium formulations a. Apoptosis was induced by culturing chp212 cells in the presence of the cyclin dependent kinase inhibitor roscovitine schutte et al. Jurkat t cells were left untreated a or treated for 4 hr with 12 m camptothecin b. 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 ability of cells to bind annexin v is still another marker considered to be specific to apoptosis. Early apoptotic cells express phosphatidylserines ps on. The annexin v apoptosis assay development of the annexin v apoptosis assay. Apc annexin vdead cell apoptosis kit with apc annexin v and. 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. In collaboration with nexins research bv, we provide the best and brightest annexin v conjugates available, including this fitc annexin v conjugate.
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. 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. Thus, the maxpar labeling protocol would not work with annexin v, unless a free sh group can be introduced to annexin v. Binding of annexin v is connected with apoptotic changes in nuclearmorphology, dna fragmentation and membrane leaflet symmetry. Anyone familiar with apoptosis interpretation n annexin vpi. Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometrybased methods. 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 imaging and flow cytometry instrument platforms. Inappropriately regulated apoptosis is implicated in disease states, such as alzheimers disease and cancer.
Annexin v7aad staining in keratinocytes springerlink. Flow cytometry of apoptosis unit 18 university of illinois. Detection of apoptotic dexamethasontreated thymocytes by annexin v staining. All annexins contain a putative pkc binding site, but only 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. Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for. Annexin v can only be used as a marker of apoptosis in cells where the plasma membrane is intact. Apoptosis can be induced by uvirradiation, as well as by numerous chemical treatments including the use of pankinase. 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. In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of. Cells were incubated with annexin v fitc in a buffer containing propidium iodide pi, cat. Annexin v binding to platelets is agonist, time and. Fitc annexin vdead cell apoptosis kit with fitc annexin v and pi, for flow cytometry catalog no. Detection of apoptosis using the bd annexin v fitc assay on the bd facsverse system.
Tacs annexin vfitc apoptosis detection kit 4830250k. However, due to membrane disintegration during necrosis, annexin v will also bind to intracellularly located ps in necrotic cells. Hingorani and others published detection of apoptosis using the bd annexin v fitc assay. Fitc annexin vdead cell apoptosis kit with fitc annexin v. Fitc annexin v apoptosis detection kit with 7aad biolegends fitc annexin v. Not for use in diagnostic or therapeutic procedures. 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. Annexin v can be used as a fluorescent probe to label apoptotic cells. Fitc annexin v apoptosis detection kit with 7aad biolegend. Annexin v staining is a common method for detecting apoptotic cells. Wash cells twice in azidefree and serumproteinfree pbs. A tool for identifying cells that are undergoing apoptosis.
Annexin v7aminoactinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. A fluorescein isothiocyanate fitc conjugated form of annexin v is used to detect ps exposing cells thereby marking apoptotic cells. Propidium iodide pi is widely used in conjunction with annexin v to. Detection of apoptosis using the bd annexin v fitc assay. Modified annexin vpropidium iodide apoptosis assay for. This apoptosis lecture explains about the structure and function of annexin v in apoptosis or cell death. Destroying the integrity of the plasma membrane will allow binding of annexin v to ps inside the cell.
They are mostly found in eukaryotic organisms animal, plant and fungi. Annexin is a common name for a group of cellular proteins. Measuring apoptosis using annexin v and flow cytometry. Bd annexin vv450 serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis manufacturer.
Annexin v or annexin a5 is a member of the annexin family of intracellular. Annexin v staining of adherent cells for flow cytometry for research use only. Using flow cytometry, titrate annexin v against the usual number of cells youll be using in your experiments, at the appropriate volume. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to phosphatidylserine ps in a calciumdependent manner. Any suggestions on the problem of too many annexin vpi. 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 assay on. Apc annexin v apoptosis detection kit with 7aad fc ruo. Aug062009 do you know a good method to detect apoptosis on adherent cells.
Pi data was ok but fitc annexin v data has a problem. 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. The relative proportion of annexin vpositive platelets varies markedly, from a few percent 4, 9 up to 70% 10. Apc annexin v apoptosis detection kit with pi biolegend. Aqueous buffered solution containing no preservative. Annexinv and apoptosis flow cytometry core facility. Annexin v binding assay as a tool to measure apoptosis in. Annexin vcy3 binding can be detected by flow cytometry ex 488 or 543 nm.
Add annexin v according to the manufacturers recommendations e. 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. There are a limited number of studies investigating annexin v binding in whole. V351 introduction apoptosis is a carefully regulated process of cell death that occurs. Microsoft powerpoint \262\323\255m\245\\\257\340\264\372\270. Annexin v, also known as pap1 or lipocortin v, is a 30 kda protein that binds to phospholipids in a calciumdependent manner 1. One should keep in mind, however, that use of the annexin v binding assay is hindered in some instances, e.
The annexin v apoptosis assay university of kansas. Annexin a5 or annexin v is a cellular protein in the annexin group. The special engineering process ensures the brighter fluorescence signal and better photo stability. Annexin 1, glucocorticoids, and the neuroendocrineimmune. Harvest cells and wash once with azide and proteinfree 1x dpbs. Fluorescent conjugates of annexin v are commonly used to identify apoptotic cells. Detection of apoptosis using annexin v and the nc3000. The principle of apoptosis detection method based on annexin v. I used bd fitc annexin v apoptosis detection kit 556547 and cell line is breast cancer cell.
Annexinv is a specific psbinding protein that can be used to detect apoptotic cells. Bd pharmingen technical data sheet pe annexin v apoptosis detection kit i product information material number. Annexin v staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. Annexin v or annexin a5 is a member of the annexin family of.
1242 758 198 143 1287 562 1620 833 210 437 39 1473 550 663 520 753 101 1440 2 243 1500 563 52 850 1524 440 114 838 302 120 132 144 1024 1345 1058 542 1156 736 956 474