RECOMBINANT
Catalog no.: AP1011AG.1 / AP1011AG.2
Name: Annexin V conjugated with Alexa-Fluor 488
Synonyms: Annexin-5, Lipocortin V, Endonexin lI, Calphobindin l (CBP-I), Placental anticoagulant protein l (PAP-I), Placental anticoagulant protein 4 (PP4), Thromboplastin inhibitor, Vascular anticoagulant-alpha (VAC-alpha), Anchorin CII
Conjugate: Alexa Fluor 488
Maximum of Absorption at 495 nm, Maximum of Emission at 519 nm
Swiss-Prot No: P48036
Gene Information: Gene Name: Anxa5, Anx5
GenelD: 11747
Source: E. coli
Purification: Affinity chromatography, SDS-PAGE
Storage buffer: PBS, 0.02% NaN3
Identity: Mass spectrometry peptide mass fingerprinting
Specificity: Binds specific to phosphatidylserine (PS) in the presence of calcium2
Applications: Annexin V is used for the detection of apoptosis in mouse and humans (not tested in other species).
Apoptosis is characterised by the loss of membrane asymmetry. In healthy cells, PS is located on the inner leaflet of the cell membrane whereas in apoptotic cells PS is found on the outer leaflet of the membrane. Hence, apoptotic events can be detected by binding of AnnnexinV-Alexa488 to the exposed PS. Since Annexin V can pass the membrane of necrotic cells, counterstaining with propidium iodide is recommended to distinguish necrotic and apoptotic cells.
Contents: 50 μl / 500 μl (5 μl are suitable for one assay)
Store at: - 20°C
Staining protocol:
1. Harvest 2x105 cells per staining and wash cells twice in cold phosphate-buffered-saline (PBS)
2. Resuspend cells in 100 μl binding buffer, add 5μl annexin V-Alexa488 to the cells and mix gently.
For detection of necrosis add additionally appropriate amounts of propidium iodide (final concentration 1 μg/ml or 1.5μM).
3. Incubate for 15min at room temperature in the dark
4. Add 400 μl binding buffer and analyse the staining in flow cytometry as soon as possible (within an hour).
Material not provided:
Binding Buffer: 10 mM Hepes pH7.4, 140 mM NaCl, 2.5 mM CaCl2
Propidium iodide: 50 μg/ml, final concentration 1 μg/ml
References:
1. Vermes l, Haanen C, Steffens-Nakken H, Reutelingsperger C (1995). A novel assay for apoptosis. Flow
cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled
Annexin V. J Immunol Methods 184, 39-51.
2. Rosenbaum S, Kreft S, Etich J, Frie C, Stermann J, Grskovic I, Frey B, Mielenz D, PoschI E, Gaipl U,
Paulsson M, Brachvogel B (2011). Identification of Novel Binding Partners (Annexins) for the Cell Death
Signal Phosphatidylserine and Definition of Their Recognition Motif. Journal of Biological Chemistry
286(7): 5708-5716.
For research use only
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