Annexin A5 (ANXA5) Protein

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Protein Name
  • anx
  • anx5
  • ANX V
  • anxa5
  • cb989
  • wu:fa98f06
  • wu:fj10f10
  • MGC89158
  • ANX5
  • ENX2
  • PP4
  • RPRGL3
  • Anx5
  • R74653
  • LC5
  • enx2
  • annexin A5
  • annexin A5b
  • Annexin A5
  • annexin 5
  • annexin A5-like
  • ANXA5
  • anxa5b
  • anxa5
  • LOC100220003
  • LOC100305020
  • LOC100342904
  • Anxa5
  • LOC100521982
Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose A bright red color fluorescent reagent for detecting of the early stages of apoptosis.
Sample Type Cell Culture
Characteristics The Annexin V-Cy3 conjugate contains the bright red fluorescent probe that can be used for detection of apoptosis by fluorescence microscopy with a rhodamine filter or by flow cytometry. Cy3 yields red fluorescence with a lambdamax emission of 570 nm.
Background During apoptosis, phosphatidylserine (PS) is translocated from the cytoplasmic face of the plasma membrane to the cell surface. Annexin V has a strong, Ca2+-dependent affinity for PS and therefore serves as a probe for detecting apoptosis.
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Pathways Apoptosis
Application Notes Detection method: Flow cytometry (Ex/EM = 543/570 nm) using FL2 channel and fluorescence microscopy
Protocol A. Incubation of cells with Annexin V-Cy3:
1. Induce apoptosis by desired methods.
2. Collect 1 x 10^5 cells by centrifugation.
3. Resuspend cells in 500 µL of 1X Annexin V Binding Buffer.
4. Add 1 µL of Annexin V-Cy3.
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 = 543 nm, Em = 570 nm) using FL2 channel. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-Cy3 (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-Cy3 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 a rhodamine filter. Cells that have bound Annexin V-Cy3 will show bright red staining on the plasma membrane.
Restrictions For Research Use only
Handling Advice Do not freeze!
Storage 4 °C
Expiry Date 12 months
Product cited in: Stubbs, Kim, Bariteau, Davis, Vempati, Minehart, Witkin, Qi, Krivtsov, Bradner, Kung, Armstrong: "Selective Inhibition of HDAC1 and HDAC2 as a Potential Therapeutic Option for B-ALL." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 21, Issue 10, pp. 2348-58, 2015 (PubMed).

Chen, Gao, Nicholas et al.: "Human herpesvirus 8 interleukin-6 contributes to primary effusion lymphoma cell viability via suppression of proapoptotic cathepsin D, a cointeraction partner of vitamin K epoxide reductase complex ..." in: Journal of virology, Vol. 88, Issue 2, pp. 1025-38, 2013 (PubMed).

Chen, Cousins, Sandford, Nicholas: "Human herpesvirus 8 viral interleukin-6 interacts with splice variant 2 of vitamin K epoxide reductase complex subunit 1." in: Journal of virology, Vol. 86, Issue 3, pp. 1577-88, 2012 (PubMed).

Guan, Xie, Leithäuser, Flossbach, Möller, Wirth, Ushmorov: "KLF4 is a tumor suppressor in B-cell non-Hodgkin lymphoma and in classic Hodgkin lymphoma." in: Blood, Vol. 116, Issue 9, pp. 1469-78, 2010 (PubMed).

Yu, Chiang, Chang, Liao, Lin: "The interferon stimulator mitochondrial antiviral signaling protein facilitates cell death by disrupting the mitochondrial membrane potential and by activating caspases." in: Journal of virology, Vol. 84, Issue 5, pp. 2421-31, 2010 (PubMed).

Kuhn, Hunsucker, Chen, Voorhees, Orlowski, Orlowski: "Targeted inhibition of the immunoproteasome is a potent strategy against models of multiple myeloma that overcomes resistance to conventional drugs and nonspecific proteasome inhibitors." in: Blood, Vol. 113, Issue 19, pp. 4667-76, 2009 (PubMed).

Chen, Sandford, Nicholas: "Intracellular signaling mechanisms and activities of human herpesvirus 8 interleukin-6." in: Journal of virology, Vol. 83, Issue 2, pp. 722-33, 2008 (PubMed).

Sato, Sanjo, Tsujimura, Ninomiya-Tsuji, Yamamoto, Kawai, Takeuchi, Akira: "TAK1 is indispensable for development of T cells and prevention of colitis by the generation of regulatory T cells." in: International immunology, Vol. 18, Issue 10, pp. 1405-11, 2006 (PubMed).

Ruiz-Vela, Opferman, Cheng, Korsmeyer: "Proapoptotic BAX and BAK control multiple initiator caspases." in: EMBO reports, Vol. 6, Issue 4, pp. 379-85, 2005 (PubMed).

Zencak, Lingbeek, Kostic, Tekaya, Tanger, Hornfeld, Jaquet, Munier, Schorderet, van Lohuizen, Arsenijevic: "Bmi1 loss produces an increase in astroglial cells and a decrease in neural stem cell population and proliferation." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 25, Issue 24, pp. 5774-83, 2005 (PubMed).

Breckenridge, Nguyen, Kuppig, Reth, Shore: "The procaspase-8 isoform, procaspase-8L, recruited to the BAP31 complex at the endoplasmic reticulum." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 99, Issue 7, pp. 4331-6, 2002 (PubMed).