Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916) contains Annexin V labeled with our proprietary green fluorescent dye iFluor 488, which allows the identification and quantitation of apoptotic cells on a single-cell basis by flow cytometry.
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- iScience 25:1050642022Small molecule SJ572946 activates BAK to initiate apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGiridhar Sekar,Geetika Singh,Xingping Qin,Cristina D Guibao,Brittany Schwam,Zintis Inde,Christy R Grace,Weixing Zhang,P Jake Slavish,Wenwei Lin,Taosheng Chen,Richard E Lee,Zoran Rankovic,Kristopher Sarosiek,Tudor MoldoveanuPubMed 36147946
Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Quantitative
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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
ANX5, ENX2, PP4, ANXA5, ANX5, ENX2, PP4, 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, PP4, PAP-I, VAC-alpha
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Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916) contains Annexin V labeled with our proprietary green fluorescent dye iFluor 488, which allows the identification and quantitation of apoptotic cells on a single-cell basis by flow cytometry.
Key facts
- Detection method
Fluorescent
- Sample types
Suspension cells, Adherent cells
- Assay type
Quantitative
- Assay Platform
Flow cytometer, Fluorescence microscope
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
Multi
- Storage information
Please refer to protocols
Notes
Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916) contains Annexin V labeled with our proprietary green fluorescent dye iFluor 488, which allows the identification and quantitation of apoptotic cells on a single-cell basis by flow cytometry. Optionally, it can be used for simultaneous staining of cells with Annexin V-iFluor488 (green fluorescence) and the nonvital dye propidium iodide (PI) (orange fluorescence) which allows the discrimination of intact cells (Annexin V-iFluor 488 negative, PI Staining Solution negative), early apoptotic (Annexin V-iFluor 488 positive, PI Staining Solution negative) and late apoptotic or necrotic cells (Annexin V-iFluor 488 positive, PI Staining Solution positive).
The iFluor 488 dye (Ex/Em = 490/525 nm) has spectral properties almost identical to those of FITC or Alexa Fluor® 488, making it convenient to be used with the common fluorescence instruments equipped with the light sources and filters for FITC, the most common fluorophore.
Apoptosis is a regulated process of cell death that occurs during embryonic development as well as maintenance of tissue homeostasis. Inappropriately regulated apoptosis is implicated in different disease states, such as neurodegeneration disease and cancer. The apoptosis program is characterized by morphologic features, including loss of plasma membrane asymmetry and attachment, condensation off the cytoplasm and nucleus, and compaction and fragmentation of the nuclear chromatin. Exposure of phosphatidylserine (PS) on the external surface of the cell membrane has been reported to occur in the early phases of apoptotic cell death, during which the cell membrane remains intact. In leukocyte apoptosis, PS on the outer surface of the cell marks the cell for recognition and phagocytosis by macrophages. The human vascular anticoagulant, annexin V, is a 35-36 kDa Ca2+ dependent phospholipid binding protein that has a high affinity for PS, and shows minimal binding to phosphatidylcholine and sphingomyelin. Changes in PS asymmetry, which can be analyzed by measuring annexin V binding to the cell membrane, are generally observed before morphological changes associated with apoptosis occurred and before membrane integrity is lost.
Alexa Fluor® 488 is the trademark of Invitrogen.
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Flow Cytometry - Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916)
The detection of binding activity of Annexin V-iFluor 488 to phosphatidylserine in Jurkat cells.
Annexin V-iFluor 488 Apoptosis Detection Kit (ab219916). Detection of phosphatidylserine (PS) exposure in Jurkat cells. Jurkat cells were left untreated (blue) or treated with 20 μM camptothecin (red) in a 37°C, 5% CO2 incubator for 4-5 hours. Cells were then incubated with Annexin V-iFluor reagent and PI for 30 minutes. The fluorescence intensitiy of Annexin V-iFluor 488 was measured with a FACSCalibur (BD Systems) flow cytometer using the FL1channel.
In live non-apoptotic cells, Annexin V-iFluor 488 conjugate detects innate apoptosis in non-induced cells, which is typically 2-6% of all cells. In apoptotic cells Annexin V-iFluor 488 conjugate binds to phosphatidylserine, which is located on the outer leaflet of the cell membrane, resulted in increased staining intensity.
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