Aposcreen Annexin V Protein : FITC (OPSB00049)

Catalog No: OPSB00049

Size:100Tests

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

• Product Format: Supplied as 100 tests in 1.0 mL of ApoScreen Annexin V Binding Buffer.
• Host: Expressed in E. coli
• Conjugation: FITC: Fluorescein Isothiocyanate
• Application: FC, IHC, ICC
• Additional Information: Description: 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
  2. Resuspend cells in cold 1X ApoScreen Annexin Binding Buffer (OOSB00003) 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 (7-AAD; OOSB00001) or ApoScreen Propidium Iodide (PI; OOSB00002)
  8. Analyze by flow cytometry immediately.
• Reconstitution and Storage: Store at 2-8C
• Concentration: Lot specific
• Characterization: To insure acceptable performance, each batch of product is tested by flow cytometry to conform to thecharacteristics of a standard reference reagent.
• Storage: Store at 2-8C.
• Application Info: Flow Cytometry: Fluorescein conjugates: 10 ul/10^6 cells. For flow cytometry, the suggested use of these reagents is in a final volume of 100 uL of ApoScreen Annexin Binding Buffer (OOSB00003).
• 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.
• Warning: Reagents 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.