Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane.
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Publications
- 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
Key facts
- Sample types
Suspension cells, Adherent cells
- Assay type
Direct
- Assay time
10m
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Target data
Function
This protein is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade.
Alternative names
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, CBP-I, PP4, PAP-I, VAC-alpha, ANXA5, ANX5, ENX2, PP4
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Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane.
Alternative names
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, CBP-I, PP4, PAP-I, VAC-alpha, ANXA5, ANX5, ENX2, PP4
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
Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane. Analysis is by flow cytometry or fluorescence microscopy.
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.
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. For more apoptosis assays, review the full set of , or the .
Supplementary info
- 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
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
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.
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