Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used to stain for phosphatidylserine present on the outside of cell membranes in apoptotic cells.
- One step staining procedure requiring minimal sample prep and just one 5 min incubation
- Includes propidium iodide (PI) dye to differentiate apoptotic cells from necrotic and non apoptotic cells
- Includes protocol for flow cytometers and fluorescence microscopes (Ex/Em 488/530 nm)
- Cited in > 500 publications
- Annexin V-FITC Reagent (ab14082) and Annexin V Binding Buffer (ab14084) also available as separate products
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- Cancers 15:2023TRPV6 Channel Is Involved in Pancreatic Ductal Adenocarcinoma Aggressiveness and Resistance to Chemotherapeutics.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGonçalo Mesquita,Aurélien Haustrate,Adriana Mihalache,Benjamin Soret,Clément Cordier,Emilie Desruelles,Erika Duval,Zoltan Pethö,Natalia Prevarskaya,Albrecht Schwab,V'yacheslav Lehen'kyiPubMed 38136316
- Nature communications 14:69002023DIAPH1-MFN2 interaction regulates mitochondria-SR/ER contact and modulates ischemic/hypoxic stress.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGautham Yepuri,Lisa M Ramirez,Gregory G Theophall,Sergei V Reverdatto,Nosirudeen Quadri,Syed Nurul Hasan,Lei Bu,Devi Thiagarajan,Robin Wilson,Raquel López Díez,Paul F Gugger,Kaamashri Mangar,Navneet Narula,Stuart D Katz,Boyan Zhou,Huilin Li,Aleksandr B Stotland,Roberta A Gottlieb,Ann Marie Schmidt,Alexander Shekhtman,Ravichandran RamasamyPubMed 37903764
- International journal of molecular sciences 24:2023Efficacy of Mesenchymal-Stromal-Cell-Derived Extracellular Vesicles in Ameliorating Cisplatin Nephrotoxicity, as Modeled Using Three-Dimensional, Gravity-Driven, Two-Layer Tubule-on-a-Chip (3D-MOTIVE Chip).Applications:Unspecified applicationReactive species:Unspecified reactive speciesEun-Jeong Kwon,Seong-Hye Hwang,Seungwan Seo,Jaesung Park,Seokwoo Park,Sejoong KimPubMed 37958709
- PloS one 18:e02903712023USP7 attenuates endoplasmic reticulum stress-induced apoptotic cell death through deubiquitination and stabilization of FBXO7.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSu Hyoun Lee,Kwang Chul ChungPubMed 37874827
- International journal of molecular sciences 24:2023Regulatory Elements Outside Established Gene Boundaries Are Required for Multilineage Differentiation of Embryonic Stem Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesVeronika V Ermakova,Nikita P Fokin,Nikolay D Aksenov,Evgeny I Bakhmet,Ekaterina V Aleksandrova,Andrey A Kuzmin,Alexey N TomilinPubMed 37895112
- Journal of cellular physiology 238:2827-28402023Mutual regulation between GDF11 and TET2 prevents senescence of mesenchymal stem cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJiaming Gao,Hao Wang,Junyan Shen,Xiaojing Liu,Xiaoqi Zhu,Ce Huang,Gongchen Li,Yao Sun,Zhongmin Liu,Yi Eve Sun,Hailiang LiuPubMed 37801347
- Frontiers in immunology 14:12440682023-induced neutrophil clustering and neutrophil extracellular trap formation depend on P2X1 purinergic receptor signaling.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGabriel Espinosa,Iván Conejeros,Lisbeth Rojas-Barón,Carlos Rodrigo Hermosilla,Anja TaubertPubMed 37854595
- Scientific reports 13:164242023Melatonin loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles reduce inflammation, inhibit apoptosis and protect rat's liver from the hazardous effects of CCL4.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlyaa Farid,Valina Michael,Gehan SafwatPubMed 37777583
- International journal of molecular sciences 24:2023The Protective Role of Oleuropein Aglycone against Pesticide-Induced Toxicity in a Human Keratinocytes Cell Model.Applications:Unspecified applicationReactive species:Unspecified reactive speciesManuela Leri,Marzia Vasarri,Emanuela Barletta,Nicola Schiavone,Maria Camilla Bergonzi,Monica Bucciantini,Donatella Degl'InnocentiPubMed 37834001
- Pharmacology research & perspectives 11:e011322023Neuroprotective effect of Kurarinone against corticosterone-induced cytotoxicity on rat hippocampal neurons by targeting BACE1 to activate P13K-AKT signaling - A potential treatment in insomnia disorder.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGuoqing Wu,Yanyan WuPubMed 37740616
Key facts
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Direct
- Assay time
- 10m
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Target data
Function
The protein expressed by gene ANXA5 is an anticoagulant protein that indirectly inhibits the thromboplastin-specific complex, a component involved in the blood coagulation cascade. This supplementary information is collated from multiple sources and compiled automatically.
Alternative names
ANX5, ENX2, PP4, ANXA5, Annexin A5, Anchorin CII, Annexin V, Annexin-5, Calphobindin I, Endonexin II, Lipocortin V, Placental anticoagulant protein 4, Placental anticoagulant protein I, Thromboplastin inhibitor, Vascular anticoagulant-alpha, CPB-I, PAP-I, VAC-alpha
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Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used to stain for phosphatidylserine present on the outside of cell membranes in apoptotic cells.
- One step staining procedure requiring minimal sample prep and just one 5 min incubation
- Includes propidium iodide (PI) dye to differentiate apoptotic cells from necrotic and non apoptotic cells
- Includes protocol for flow cytometers and fluorescence microscopes (Ex/Em 488/530 nm)
- Cited in > 500 publications
- Annexin V-FITC Reagent (ab14082) and Annexin V Binding Buffer (ab14084) also available as separate products
Key facts
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Direct
- Assay time
- 10m
- Assay Platform
- Flow cytometer, Fluorescence microscope
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
How the assay works
Annexin V-FITC Apoptosis Staining / Detection kit uses FITC labeled protein Annexin V that binds to the phosphatidylserine molecules on the apoptotic cell membrane, which is normally located on the inner side of a healthy cell, allowing the identification and quantification of early apoptotic cells using flow cytometry or a fluorescent microscope.
The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
The Annexin V-FITC reagent contained in the kit is also available as Annexin V-FITC reagent Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X) ab14082.
Annexin V-FITC Apoptosis Staining / Detection Kit protocol summary:
- Collect cells
- Resuspend cells in binding buffer.
- Add Annexin V-FITC and Propidium Iodide and incubate.
- Analyze by flow cytometry to quantify the percentage of apoptotic cells or by fluorescent microscope
Related and recommended products
See other alternative kits to quantify apoptosis using Annexin V with different labels:
- Annexin V-Cy5 Apoptosis Staining / Detection Kit Annexin V-Cy5 Apoptosis Staining / Detection Kit ab14150
- Annexin V-PE Apoptosis Staining / Detection Kit (with dead cell stain) Annexin V-PE Apoptosis Staining / Detection Kit (with dead cell stain) ab273274
- Annexin V-iFluor 350 Apoptosis Staining / Detection Reagent Annexin V-iFluor 350 Apoptosis Staining / Detection Reagent ab219903
- Annexin V-iFluor 488 Apoptosis Staining / Detection Reagent Annexin V-iFluor 488 Apoptosis Staining / Detection Reagent ab219904
Soon after initiating apoptosis, cells translocate membrane phosphatidylserine molecules from the inner face of the plasma membrane to the cell surface. Phosphatidylserine on the cell surface is detected by staining with a fluorescent conjugate of Annexin V, a protein that has a high affinity for phosphatidylserine.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Annexin V also known as ANXA5 is a protein with a mass of approximately 35-36 kDa. This protein belongs to the annexin family which includes proteins capable of binding to phospholipids in a calcium-dependent manner. Annexin V is mostly expressed in placental tissue though it can be found in various other tissues like the vascular endothelium. Researchers often utilize annexin V for staining dead cells or identifying apoptosis using assays such as annexin V FITC or 7-AAD staining. It forms part of the annexin V binding buffer which enhances its binding affinity.
- Biological function summary
Annexin V has an important role in cell membrane dynamics and signaling. The protein does not function as part of a larger complex but its ability to bind negatively charged phospholipids like phosphatidylserine makes it important in apoptosis. During apoptosis phosphatidylserine translocates to the outer leaflet of the plasma membrane where annexin V marks these apoptotic cells. This property enables annexin V-based assays to detect early stages of apoptosis.
- Pathways
Annexin V involvement is seen in pathways associated with blood coagulation and apoptosis. In the apoptosis pathway annexin V interacts with apoptotic markers to identify dying cells. It relates to proteins such as caspases which are key executors of apoptosis. In coagulation annexin V has been shown to interact with prothrombinase complex and can affect calcium ion concentration thereby influencing blood clotting processes.
- Associated diseases and disorders
Annexin V plays a role in antiphospholipid syndrome and cardiovascular diseases. Annexin V has been researched for its connection to antiphospholipid antibodies where it affects normal annexin V function impacting clot formation. In cardiovascular diseases annexin V can interact with proteins such as thrombomodulin influencing thrombotic and atherosclerotic pathways. Understanding these interactions helps researchers develop potential therapeutic approaches for related conditions.
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4 product images
Functional Studies - Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
Annexin V-FITC/ PI staining of AG06173 primary fibroblasts.
105 cells were used for analysis. Resuspended cells were incubated with Annexin V-FITC for 15 min in the dark. Propidium iodide (Propidium Iodide ab14083) was used as a counterstain to discriminate necrotic/ dead cells from apoptotic cells. Left: negative control - AG6173 untreated cells. Right: positive control - AG6173 cells irradiated at 10 Gy.
Image courtesy of S. Khoronenkova PhD, Gray Institute for Radiation Oncology and Biology, University of Oxford, Oxford, UK.
Alpay et al., PLos One, 9(19), Fig 5B. Doi:10.1371/journal.pone.0105526 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Flow Cytometry - Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
Flow cytometery analysis of treated HeLa cells for 48 hours.
HeLa cells were harvested with trypsinization together with floating non-viable cells. Cells were washed with PBS and suspended in sodium citrate buffer 20 minutes prior to analysis. HeLa cells were treated with Mitoxantrone (MX) and MX +Imatinib for 48 hours. The samples were then stained with Annexin V-FITC Apoptosis Staining/Detection kit (ab14085). A FACSCalibur flow cytometer was used for cell cycle analysis.
This is a modified version of the original image
Image courtesy of an anonymous customer reviewImmunocytochemistry/ Immunofluorescence - Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
Apoptosis in Mouse Cortical Collecting Duct Cells.
ab14085 was used to determine minor levels of apoptosis (using both the Annexin V-FITC and PI) in mouse cortical collecting duct cellss (mCCDs). mCCD cells were incubated with serum free medium for 48h. The green label on the plasma membrane (Annexin V-FITC) and the absence of nuclear red (PI) staining indicates apoptosis rather than necrosis. Fluorescent microsocpy ws used to analyse the cells.
Guo W et al., PLoS One, 8(8). Fig7b, doi: 10.1371/journal.pone.0070910 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
Analysis of apoptosis in prostate cancer cells following treatment with CTA095.
PC3 cells were seeded at 106 cells/ml and incubated overnight and then treated with CTA095 at various concentrations for 24hours. Apoptosis was then analyzed using Annexin-V FITC apoptosis detection kit (ab14085).
This is a modified version of the original image
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