Annexin A5 (ANXA5) protein (PE,Cy5)

Name: Annexin A5 (ANXA5)

Synonyme: PP4, ANX5, ENX2, Anx5, R74653, LC5, MGC93655, ANXA5, MGC126971, anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158

Konjugat: PE,Cy5

Applikation: Durchflusszytometrie (FACS), Fluoreszenzmikroskopie (FM)

Produktnummer: ABIN412020

Produktbeschreibung

Beschreibung: Annexin V-PE-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, Ca 2+ -dependent affinity for PS and therefore serves as a probe for detecting apoptosis. The one-step staining procedure takes only 10 minutes. In addition, the assay can be directly performed on live cells. Annexin V-PE-Cy5 stained cells can be detected by flow cytometry or fluorescence microscopy.

Anwendungen

Protokoll: A. Incubation of cells with Annexin V-PE: 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-PE-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 Annexin V-PE-Cy5 binding by flow cytometry (Ex = 488 nm, Em = 670 nm). For analyzing adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-PE-Cy5 (A.3-5). C. Detection by Fluorescence Microscopy 1. Place the cell suspension from Step A.5 on a glass slide. Cover the cells with a glass coverslip. For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on glass slide and visualize cells. The cells can also be washed and fixed in 2% paraformaldehyde before visualization. (Cells must be incubated with Annexin V-PE-Cy5 before fixation since 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 a PE-Cy5 filter. Cells which have bound Annexin V-PE-Cy5 will show bright purple staining in the plasma membrane. STORAGE AND STABILITY: Store at 4 °C. Stable for one year under proper storage conditions.

Lagerung: Store at 4 °C. Do not freeze. SHELF LIFE: 1 year under proper storage conditions

Beschränkungen: Nur für Forschungszwecke einsetzbar