Annexin v staining materials 12 x 75 mm roundbottom tubes 1x pbs annexin v apoptosis detection kit cat. Specificity the annexin vfitc kit is an apoptosis detection kit based on the binding properties of annexin v to phosphatidylserine ps and on the dnaintercalating capabilities of. Fitc annexin v apoptosis detection kit ii bd biosciencesus. Inappropriately regulated apoptosis is implicated in disease states, such as alzheimers disease and. 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 v and 7aad positive, in of itself, reveals less information about the process by which the cells underwent their demise. Scatter plots and histograms were obtained from the nucleoview nc3000 software. These studies report binding of annexin v to platelets after stimulation with. An additional control that may be performed includes preincubation of cell samples with recombinant unconjugated annexin v, which is included as part of the annexin vfitc apoptosis detection kit ii cat.
This antibody is not known or predicted to crossreact with other annexin family members. Annexins are traditionally thought of as calciumdependent phospholipidbinding proteins, but recent work suggests a more complex set of functions. Prior to staining cells, an appropriate quantity of a 1x working solution should be made by diluting the 10x concentrate 1. The annexin v binding buffer is a 10x concentrate composed of a 0. Bd phospholipidbinding protein, apc annexin v apc annexin.
When cells are ethidium bromide negative, they can still be annexin v positive. Ive been using bds cell cycle test plus kit and i am a beginner in this kind of experiment. Also, damage to cell membranes from dissection or sectioning of tissues could result in. Measuring apoptosis using annexin v and flow cytometry. Annexin v staining of early chicken and mammalian embryos in culture has been reported in the scientific literature. Bd accuri c6 flow cytometer simplify the detection of apoptosis at different stages. Although their functions are still not clearly defined, several members of the annexin family have been implicated in membranerelated events along exocytotic and. Apoptosis is a programmed, physiological mode of cell death, that plays an important role during embryonic development, the maintenance of tissue homeostasis and the deletion of aberrant cells kerr and harmon, 1991. Annexin ii bd biosciences 610069 product information. Genotoxic agents promote the nuclear accumulation of annexin a2. Annexin a1 and glucocorticoids as effectors of the. Microsoft powerpoint \262\323\255m\245\\\257\340\264\372\270. The reagents may then be diluted in complete medium. The relative proportion of annexin vpositive platelets varies markedly, from a few percent 4, 9 up to 70% 10.
I use an annexin v that is conjugated with fitc fluorescein isothiocyanate. For example, association of annexin vi with membranes was found to induce extensive clustering of acidic phospholipids in a calciumdependent manner bazzi and nelsestuen, 1991a. This may also be excited with a wavelength of 488nm light and has an emission peak at 521nm. Stain them with annexin v as determined in the previous step, using 0, 0. Bd pharmingen technical data sheet annexin vfitc apoptosis detection kit i product information catalog number. The annexin v pi protocol is a commonly used approach for studying apoptotic cells3. 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.
All annexins contain a putative pkc binding site, but only annexin v has been identified as an inhibitor of this pathway 2. Bd, the bd logo and all other trademarks are property of becton, dickinson and company. Telecharger des films, series, jeux, logiciels, albums et magazines gratuitement ragnar 8 integrales fr hd pdf cbr bd multi categories. They are mostly found in eukaryotic organisms animal, plant and fungi. Annexin a2 d11g2 rabbit mab recognizes endogenous levels of total annexin a2 protein. Detection of apoptotic dexamethasonetreated thymocytes by annexin v staining. This serves to block annexin vfitc binding sites and thus demonstrates the specificity of annexin vfitc.
Telecharger ebook gratuit, magazines et journaux en epub, pdf. A good positive control is vital when getting started. Annexin v labeled with a fluorophore or biotin can identify apoptotic cells by binding to ps exposed on the outer leaflet. Modified annexin vpropidium iodide apoptosis assay for. Bd pharmingen fitc annexin v apoptosis detection kit. Annexin a1 subcellular expression in laryngeal squamous. Annexin a6 is a protein that in humans is encoded by the anxa6 gene. 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. Add annexin v and calcium only 10 minutes before acquisition. Modified annexin vpropidium iodide apoptosis assay for accurate.
Detection of apoptotic dexamethasontreated thymocytes by annexin v staining. Other applications were tested at bd biosciences pharmingen during antibody development only or reported in the literature. The combination of annexin vfitc and propidium iodide allows for the. Thymocytes were left untrated left or treated with 100 nm dexamethasone for 15. 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. Apoptosis is an active process governed by many genes that either promote or block cellular breakdown from within. Ps is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and ps translocates to the external leaflet.
Harvest cells and wash once with azide and proteinfree 1x dpbs. Depletion of annexin a1 or a2 alters the retrograde transport of shiga toxin but not ricin, without affecting toxin binding or internalization. Annexin a5 or annexin v is a cellular protein in the annexin group. Annexin a1 protein has an apparent relative molecular mass of 40 kda with phospholipase a2 inhibitory activity. Simple one step assay kit to detect apoptosis within 10 min. Comparative study of human and mouse postsynaptic proteomes finds high compositional. Annexin v, also known as pap1 or lipocortin v, is a 30 kda protein that binds to phospholipids in a calciumdependent manner 1. Pi is used more often than other nuclear stains because it. Telecharger votre livre asterix obelix et compagnie n23 au format pdf ou epub. Apc annexin v is routinely tested by flow cytometric analysis. There are a limited number of studies investigating annexin v binding in whole blood 11, 12.
Annexin v staining is a common method for detecting apoptotic cells. Annexin a11 is one of the oldest vertebrate annexins that has a crucial role in sarcoidosis pathogenesis. 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. Annexinvfluos can be directly detected in facs analysis and immunochemistry without a secondary detection system. The kit contains recombinant annexin v conjugated to rphycoerythrin rpe. Detection of apoptosis using annexin v and the nc3000. Early apoptotic cells express phosphatidylserines ps on the outer leaflet of the plasma membrane. Bd phospholipidbinding protein, apc annexin v has high affinity for ps, and binds to cells with exposed ps. 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 of the cells to the 36 kda annexin v protein.
Annexin vfitc kit detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin v conjugated to greenfluorescent fluorescein isothiocyante fitc dye. However some annexins annexin a1, annexin a2, and annexin a5 have also been found outside the cellular environment, for example, in blood. Jurkat cells were grown in the absence upper row or in the presence lower row of camptothecin cpt. Even uvc if you have a stratalinker handy, can be used. Annexin v binding to platelets is agonist, time and. In flow cytometry, annexin v is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.
Fluorescent conjugates of annexin v are commonly used to identify apoptotic cells. Dilute apoptosis reagents in desired medium formulations a. More than a thousand proteins of the annexin superfamily have been identified in major eukaryotic phyla, but annexins are absent from yeasts and prokaryotes. Annexin v binding buffer, 10x concentrate bd biosciences. Only cells that stain double positive demonstrate chromatin condensation. Annexin vfitc kit system for detection of apoptosis pn im2375 20 tests page 1 of 2 im2375 20110926 for research use only. Bd pharmingen technical data sheet fitc annexin v apoptosis detection kit ii product information material number. Tacs annexin vfitc apoptosis detection kit 4830250k.
Le telechargement gratuit pour windows ou mac os prendre moins dune minute pour installer via une connexion a large bande. 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. Anyone familiar with apoptosis interpretation n annexin vpi. Binding of annexin v is connected with apoptotic changes in nuclearmorphology, dna fragmentation and membrane leaflet symmetry.
Annexin v binding assay as a tool to measure apoptosis in. Cells which still mantain membrane integrity for longer incubation times may become positive for pi since this dye will enter intact cells although very slowly. Wash cells twice in azidefree and serumproteinfree pbs. Apc annexin v apoptosis detection kit with pi biolegends apc annexin v. Annexin v7aad staining in keratinocytes springerlink. Bd biosciences technical resources page 3 february 20 annexin v staining of adherent cells for flow cytometry for research use only. Lactadherin detects early phosphatidylserine exposure on. The combination of annexin vfitc and propidium iodide allows for the distinction. Annexin v or annexin a5 is a member of the annexin family of intracellular proteins that binds to phosphatidylserine ps in a calciumdependent manner. The annexin vpi protocol presented here is a modified version of conventional protocols and takes into account the presence of rna in the cytoplasm which also has high affinity for pi. Annexin v or annexin a5 is a member of the annexin family of. After taking the cells from the culture keep them on ice at all times. Annexin vfitc kit system for detection of apoptosis.
In this opinion article, the effector function of annexin a1 in the antiinflammatory action of glucocorticoids is discussed, and a model to address the opposing effects of glucocorticoids on the. The unique annexin core domain is made up of four similar repeats approximately 70. Annexinlipid interactions are probably of the same kind as those described for pkc, if results of studies on annexin vi can be applied generally to the whole family. Not for use in diagnostic or therapeutic procedures. Annexin v7aminoactinomycin staining is a convenient way to discriminate early apoptosis from late apoptosis and necrosis. Apc annexin v apoptosis detection kit with pi biolegend. Annexin is a common name for a group of cellular proteins. I actually make the annexinbuffer and pi solution myself as they are easy to make and it saves money. The mechanism of effect in sarcoidosis granuloma cells may be due to alterations in apoptosis. V241 introduction apoptosis is a carefully regulated process of cell death that occurs as a normal part of development. The values are shown in percentage of diploid and tetraploid or aneuploid cells on g1, s and g2m. Annexin v is marketed with several different flurophores. Hingorani and others published detection of apoptosis using the bd annexin v fitc.