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Annexin A5 (ANXA5) protein (EGFP)
|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)
|7 references available|
|Price||324.50 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 2 to 3 Business Days|
|Description||The bright and photo-stable reagent for detecting 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. The annexin V-EGFP fusion is brighter and photo-stable, ideal for detecting apoptosis by fluorescence microscopy with a FITC filter or by flow cytometry.|
|Protocol||A. Incubation of cells with Annexin V-EGFP 1. Induce apoptosis by desired method. 2. Collect 1-5 x 10 5 cells by centrifugation. 3. Resuspend cells in 500 µl of 1X Binding Buffer (Cat.# 1035-100). 4. Add 1 µl of Annexin V-EGFP and 1 µl of propidium iodide (PI, Cat.# 1056-1) 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-EGFP binding by flow cytometry (Ex = 488 nm, Em = 530 nm) using FITC signal detector (usually FL1) and PI staining by the phycoerythrin emission signal detector (usually FL2). For adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-EGFP (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% formaldehyde before visualization.|
|Application Notes||Note: Cells must be incubated with Annexin V-EGFP 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 dual filter set for FITC & rhodamine. Cells which have bound Annexin V-EGFP will show green staining in the plasma membrane. Cells which have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (EGFP) on the cell surface (plasma membrane).|
|Expiry Date||12 months|
|Restrictions||For Research Use only|
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