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Annexin A5 (ANXA5) protein (Cy5)
|Synonyms||PP4, ANX5, ENX2, Anx5, R74653, LC5, MGC93655, ANXA5, MGC126971, anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158|
Flow Cytometry (FACS), Fluorescence Microscopy (FM)
|9 references available|
|Price||1,094.50 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 2 to 3 Business Days|
|Description||Annexin V-Cy5 is a bright fluorescent reagent for detecting of the early stages of apoptosis. During apoptosis, phosphatidylserine (PS) is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca²ᶧ-dependent affinity for PS and therefore serves as a probe for detecting apoptosis. Cy5 fluorescent dye produces an intense signal in the far-red region of the spectrum and therefore it is very useful for multiple labeling of cells with green and red colored fluorescent probes. Cy5 yields fluorescence with a λmax emission of 670 nm.|
|Protocol||A. Incubation of cells with Annexin V-Cy5: 1. Induce apoptosis by desired methods. 2. Collect 1 x 10 5 cells by centrifugation. 3. Resuspend cells in 500 μ l of 1X Annexin V Binding Buffer (Cat.#1035-100). 4. Add 1 μ l of Annexin V-Cy5. 5. Incubate at room temperature for 5 min in the dark. Proceed to B or C below depending on method of analysis. B. Quantification by Flow Cytometry: Analyze cells by flow cytometry (Ex = 649 nm, Em = 670 nm) using Helium-Neon Laser. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-Cy5 (A.3-5). C. Detection by Fluorescence Microscopy: 1. Place the cell suspension from Step A.5 on a glass slide, and cover with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on a glass slide and visualize cells. The cells can also be washed with 1X Annexin V Binding Buffer and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-Cy5 before fixation because any cell membrane disruption can cause nonspecific binding of annexin V to PS on the inner surface of the cell membrane.) 2. Observe the cells under a fluorescence microscope using Cy5 filter or a FITC/Cy3/Cy5 triple band filter set (Chroma Technology) if you perform triple labeling with these dyes, or detect cells using CCD camera. Cells that have bound Annexin V-Cy5 will show bright red-blue staining on the plasma membrane.|
|Expiry Date||12 months|
|Restrictions||For Research Use only|
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