Catalog No: OPSB00050
Price: $199.00
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Datasheets/ManualsPrintable datasheet for OPSB00050
Product Info
HostExpressed in E. coli
ApplicationFC, IHC, ICC
Additional InformationDescription: ApoScreen Annexin V is a recombinant protein that is expressed in E. coli. A number of changes in cell structure occur during apoptosis. One major change is the loss of membrane phospholipid asymmetry, with translocation of phosphatidylserine (PS) from the inner leaflet of the phospholipid bilayer to the external surface. Annexin V, a member of the annexin family of calcium-dependent phospholipid binding proteins, has a high affinity for phosphatidylserine (PS)-containing phospholipid bilayers. Conjugated Annexin V can be used to monitor changes in cell membrane phospholipid asymmetry, thereby providing a convenient tool for detection of apoptotic cells.
::Suggested Staining Protocol:
1. Wash cells twice in cold PBS; remove the PBS from the cell pellet after the second wash.
2. Resuspend cells in cold binding buffer (e.g. 10 mM HEPES pH 7.4, 140 mM NaCl, 2.5 mM CaCl2, 0.1% BSA) to a concentration of 1 x 106 to 1 x 107 cells/mL.
3. Add 100 uL of cells (1 x 105 to 1 x 106) to each of four labeled tubes.
4. Add 10 uL of labeled annexin V to tube #2 and tube #4.
5. Gently vortex each tube and incubate for 15 minutes on ice, protected from light.
6. Without washing, add 380 uL of cold 1X binding buffer to each tube. Add 10 uL of PI (from a 50 ug/ml stock solution) to tube #3 and tube #4.
7. Immediately analyze by flow cytometry. Note: the 4-tube protocol above is for initially adjusting settings on the flow cytometer; subsequent assays should basically require the protocol described for 'tube #4,' i.e. cells stained with both labeled annexin V and PI.
The Biotin (BIOT) conjugate is supplied as 100 tests in 1.0 mL of 1X Binding Buffer. Store at 2-8C.
Reconstitution and StorageStore at 2-8C
DilutionFlow Cytometry: Biotin conjugate
10 ul/106 cells
ConcentrationLot specific
CharacterizationTo insure acceptable performance, each batch of product is tested by flow cytometry to conform to thecharacteristics of a standard reference reagent.
Storage Annexin V- BIOT: Store at 2- 8 C
Application InfoBlocking Annexin V-conjugates, Flow cytometry, Fluorescence microscopy
Other Applications DataSince applications vary, you should determine the optimum workingamount of the product that is appropriate for your specific need.
Reference1. Fadok, V.A. et al. 1992. J. Immunol. 148:22072. Tait, J.F., et al. 1989. J. Biol. Chem. 264:7944
3. Andree, H.A., et al. 1990. J. Biol. Chem. 265:4923
4. Thiagarajan, P., et al. 1990. J. Biol. Chem. 265:17420
5. Dachary, P.J., et al. 1993. Blood 81:2554
6. Koopman, G., et al. 1994. Blood 84:1415
7. Martin, S.J., et al. 1995. J. Exp. Med. 182:1545
8. Wood, B.L., et al. 1996. Blood 88:1873.
WarningReagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin,eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash withcopious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxichydrazoic acid under acidic conditions. Dilute azide- containing compounds in running water before discardingto avoid accumulation of potentially explosive deposits in lead or copper plumbing.
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