Aposcreen Annexin V Protein : UNLB (OPSB00048)

Catalog No: OPSB00048

Size:100UG

Amount Used: 10 uL/106 cells Two-color flow cytometry analysis of untreated (left panel) and irradiated (right panel) BALB/c thymocytes stained with annexin V-FITC and propidium iodide. Early apoptotic cells are located in the lower right quadrant of the two displays.

Product Info

• Host: Expressed in E. coli
• Conjugation: Unconjugated
• Application: FC, IHC, ICC
• Reconstitution and Storage: Store material at -20C prior to reconstitution. Reconstitute with 1.0 mL of dH₂O. Store reconstituted material at 2-8C.
• Characterization: To insure acceptable performance, each batch of product is tested by flow cytometry to conform to thecharacteristics of a standard reference reagent.
• Procedure: 1.Wash cells twice in cold PBS.
  2. Resuspend cells in cold 1X ApoScreen Annexin Binding Buffer to a concentration of 1 x 10⁶ cells/mL.
  3. Add 100 uL of cell suspension to 5 mL polystyrene round bottom tubes.
  4. Add 10 uL of conjugated ApoScreen Annexin V.
  5. Gently vortex each tube and incubate for 15 minutes at 2-8C and protected from light.
  6. Without washing, add ~380 uL of cold 1X ApoScreen Annexin Binding Buffer to each tube.
  7. Add 10 uL of ApoScreen 7-Aminoactinomycin D or ApoScreen Propidium Iodide.
  8. Analyze by flow cytometry immediately.
• Application Info: Blocking Annexin V-conjugates, Flow cytometry, Fluorescence microscopy
• Other Applications Data: Since applications vary, you should determine the optimum workingamount of the product that is appropriate for your specific need.
• Reference: 1. 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.

Target Info

• Description of Target: 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.