Annexin A5 (ANXA5) protein (Cy5)

Name: Annexin A5 (ANXA5)

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

Host: Please enquire

Reactivity: Please enquire

Conjugate: Cy5

Application: Flow Cytometry (FACS), Fluorescence Microscopy (FM)

Catalog no.: ABIN412016

Additional Information

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, Ca 2+ -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 λ max emission of 670 nm.

Application Details

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.

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

Restrictions: For Research Use only

Publications

Product: Smith, Waelde, Horwitz et al.: "Evaluation of the role of phosphatidylserine translocase activity in ABCA1-mediated lipid efflux." in: The Journal of biological chemistry, Vol. 277, Issue 20, pp. 17797-803, 2002 (PubMed).

Wang, Hasegawa, Imamura et al.: "PYNOD, a novel Apaf-1/CED4-like protein is an inhibitor of ASC and caspase-1." in: International immunology, Vol. 16, Issue 6, pp. 777-86, 2004 (PubMed).

Zhang, Soboloff, Zhu et al.: "Inhibition of Ca2+ influx is required for mitochondrial reactive oxygen species-induced endoplasmic reticulum Ca2+ depletion and cell death in leukemia cells." in: Molecular pharmacology, Vol. 70, Issue 4, pp. 1424-34, 2006 (PubMed).

Wen, Leung, Huang et al.: "Survivin is not required for the endomitotic cell cycle of megakaryocytes." in: Blood, Vol. 114, Issue 1, pp. 153-6, 2009 (PubMed).

Hole, Pearn, Tonks et al.: "Ras-induced reactive oxygen species promote growth factor-independent proliferation in human CD34+ hematopoietic progenitor cells." in: Blood, Vol. 115, Issue 6, pp. 1238-46, 2010 (PubMed).

Keerthivasan, Small, Liu et al.: "Vesicle trafficking plays a novel role in erythroblast enucleation." in: Blood, Vol. 116, Issue 17, pp. 3331-40, 2010 (PubMed).

Corcoran, Dias-Santagata, Bergethon et al.: "BRAF gene amplification can promote acquired resistance to MEK inhibitors in cancer cells harboring the BRAF V600E mutation." in: Science signaling, Vol. 3, Issue 149, pp. ra84, 2010 (PubMed).

Corcoran, Contino, Deshpande et al.: "STAT3 plays a critical role in KRAS-induced pancreatic tumorigenesis." in: Cancer research, Vol. 71, Issue 14, pp. 5020-9, 2011 (PubMed).

Greco, Quintiliani, Santucci et al.: "Janus-faced liposomes enhance antimicrobial innate immune response in Mycobacterium tuberculosis infection." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, Issue 21, pp. E1360-8, 2012 (PubMed).