Annexin v binding to platelets is agonist, time and. Bd biosciences 2350 qume drive san jose, ca 951 detection of apoptosis on the bd facsverse system a streamlined approach to application support. V fitc apoptosis detection kit provides a set of reagents for the detection of apoptosis using flow cytometry. Apoptosis will be detected by initially staining the cells with annexin v and propidium iodide solution followed by. Detection of apoptosis using the bd annexin v fitc assay. Posttranslational modifications of annexin a2 are linked to its association with perinuclear nonpolysomal mrnp complexes. Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometrybased methods. Modified annexin vpropidium iodide apoptosis assay for. Bd pharmingen will not be held responsible for patent infringement or other violations that may occur with the. Cells that stain negative for both fitc annexin v and pi are alive and not undergoing measurable apoptosis. Propidium iodide pi is widely used in conjunction with annexin v to determine if cells are viable, apoptotic, or necrotic through differences in plasma membrane integrity and permeability 1,2. Bd phospholipidbinding protein, apc annexin v apc annexin.
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. In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of. Annexin v and 7aad negative viable, or no measurable apoptosis, to pe annexin v positive and 7aad negative early apoptosis, membrane integrity is present and finally to pe annexin v and 7aad positive end stage apoptosis and death. These cells will stain with annexin v but not with viability dyes, thus distinguishing cells in early apoptosis. Fitc annexin v apoptosis detection kit with pi fc ruo. The relative proportion of annexin v positive platelets varies markedly, from a few percent 4, 9 up to 70% 10. Fitc annexin v apoptosis detection kit with 7aad fc ruo. Bd lsr ii flow cytometer fluidigm cytofmass cytometer mission to ensure that university of maryland investigators have access to flow cytometry and mass cytometry services for their research. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to. Fitc annexin vdead cell apoptosis kit with fitc 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. Reading this material and answering the questions at the end of each section will enhance your handson training experience during operator training at bd. Bd pharmingen technical data sheet annexin v fitc apoptosis detection kit ii product information catalog number. To understand the principles of flow cytometry and system.
Flow cytometry of apoptosis unit 18 university of illinois. Annexin vcf blue 7aad apoptosis staining detection kit. Annexin v cf blue 7aad apoptosis staining detection kit ab214663 contains annexin v labeled with cf blue, which allows the identification and quantitation of apoptotic cells on a singlecell basis by. Cells that stain positive for both pe annexin v and 7aad are either in the end stage of apoptosis, are undergoing necrosis, or are already dead. Pdf detection of apoptosis using the bd annexin v fitc. Pe,annexin,v,apoptosis,detection,kit,i,ruo 559763 bd.
Annexin ii bd biosciences 610069 product information. I used annexin v kit from bd and followed the protocol in the kit. Methods for utilizing annexin v binding on adherent cells i. Bd phospholipidbinding protein, apc annexin v has high affinity for ps, and binds to cells with exposed ps. 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. Bd pharmingen technical data sheet annexin v pe apoptosis detection kit i product information catalog number.
Fixable blue dapi sytox blue hoechst 33342 vybrant dyecycle violet fxcycle violet dapi annexin v, alexa fluor 350 annexin v. Production of recombinant human annexin v by fedbatch cultivation. Annexin v binds to phosphatidylserine, which migrates to the outer plasma membrane in apoptosis. Annexin a5 or annexin v is a cellular protein in the annexin group. Cells that stain negative for both pe annexin v and 7aad are alive and not undergoing measurable apoptosis. Ps interaction, it is critical to avoid buffers containing edta or other calcium chelators during annexin v experiments. Detection of apoptosis on the bd facsverse system a. Annexin v pe and 7aad are either in the end stage of apoptosis, are undergoing necrosis, or are already dead.
Cells that stain positive for both fitc annexin v and pi are either in the end stage of apoptosis, are undergoing necrosis, or are already dead. This format retains its high affinity for ps and thus serves as a sensitive probe for flow cytometric. In order to make that calcuation using becton dickinson flow cytometers, the total volume of cell sample fluid passing through the instrument during data acquisition must be determined. The fitc annexin v dead cell apoptosis kit with fitc annexin v and pi for flow cytometry provides a rapid and convenient assay for apoptosis. Annexin v may be conjugated to fluorochromes including fitc. Protocol for apoptosis assay by flow cytometry using. Here we describe only the usage of the annexin v pi labeling solutions from bd.
The becton dickinson flow cytometers do not calculate absolute cell counts total number of cells per sample. Staining precedes the loss of membrane integrity which accompanies the latest stages of cell death resulting from. This assay is used to count the number of cells that have undergone apoptosis. Apc annexin v apoptosis detection kit with pi biolegend. The bd annexin v fitc assay shown illustrates the detection of camptothecininduced apoptosis in jurkat cells using the bd pharmingen annexin v. Bd annexin v v450 serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. From the laboratory for cellular and molecular immunology, headed by dror mevorach, md, at the hadassahhebrew university medical center, israel. Pacific blue annexin v apoptosis detection kit with 7aad. Bd biosciences provides flow cytometers, reagents, tools, and a wide range of services to support the work of researchers and clinicians who understand disease and improve care. Thermo fisher scientific offers highquality fluorescent annexin v conjugates as standalone reagents and in a variety of kits for. What is the general consensus on flow cytometry about.
Annexin v staining protocol general annexin v staining procedure solutions. Fitc,annexin,v,apoptosis,detection,kit,ruo 556547 bd. Annexin v may be conjugated to fluorochromes including cy5. Annexin v staining is a common method for detecting apoptotic cells. Therefore, we compared the results obtained by staining with 1, 5, 10, and 20. Annexin v is a calciumdependent phospholipidbinding protein that has a high affinity for ps, and is useful for identifying apoptotic cells with exposed ps. Buy two bd horizon brilliant reagents and get a vial of brilliant stain buffer at no. Annexin v will also bind to intracellularly located ps in necrotic cells. Dead cell apoptosis kit with annexin v fitc and pi, for. Propidium iodide pi is a standard flow cytometric viability probe and is used to distinguish viable from. Kit product description 556547 bd pharmingen fitc annexin v apoptosis detection kit i kit components kit components description 51 66211e bd. The kit contains recombinant annexin v conjugated to fluorescein fitc annexin v. Flow cytometric analysis of apoptotic cells stained with both fitclabeled annexin v and pi using the bd biosciences kit or a homemade kit.
There are a limited number of studies investigating annexin v binding in whole blood 11, 12. One should keep in mind, however, that use of the annexin v binding assay is. The ability of cells to bind annexin v is still another marker considered to be specific to apoptosis. Shop online for a wide selection of bd annexin v fitc serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. Becton, dickinson and company bd, franklin lakes, nj 07417 usa. This product detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin v conjugated to greenfluorescent fitc dye and dead cells using propidium iodide pi. Bd biosciences provides flow cytometers, reagents, tools, and a wide range of.
Pe apoptosis detection kit i contains apoptotic program characterized by certain morphologic features, including loss of plasma membrane asymmetry and attachment, condensation of the cytoplasm and nucleus, and internucleosomal cleavage of dna. Pe apoptosis detection kit i contains apoptotic program characterized by certain morphologic features, including loss of plasma membrane asymmetry and attachment, condensation. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v apoptosis. The cells become reactive with annexin v prior to the loss of the plasma membranes ability to exclude cationic dyes such as pi or 7aminoactinomycin d 7aad.
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