Annexin A5 (ANXA5) Protein

Details for Product No. ABIN412007
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Protein Name
Synonyms anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158, ANX5, ENX2, PP4, RPRGL3, Anx5, R74653, LC5, enx2
(35), (9), (4), (4), (4), (4), (2)
(38), (9), (7), (2), (2)
Flow Cytometry (FACS), Fluorescence Microscopy (FM)
Pubmed 21 references available
Quantity 1000 tests
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Purpose Reagent for detecting early stages of apoptosis.
Sample Type Cell Culture
Characteristics The Annexin V-FITC conjugate can be used for detection of apoptosis by fluorescence microscopy or by flow cytometry.
Alternative Name Annexin V
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 is used as a probe for detecting apoptosis.
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Application Notes Detection method: Flow cytometry (Ex/Em = 488/530 nm) using FL1 channel and fluorescence microscopy
Protocol A. Incubation of cells with Annexin V-FITC:
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 (ABIN412353).
4. Add 1 µL of Annexin V-FITC and 1 µL Propidium Iodide (ABIN412371).
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 = 488 nm, Em = 530 nm) using FL1 channel for detecting Annexin v-FITC staining and FL2 channel for detetcing PI staining. For adherent cells, trypsinize and gently wash cells with serum-containing medium before incubation with Annexin V-FITC (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 % formaldehyebefore visualization. (Cells must be incubated with Annexin V-FITC 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 dual filter set for FITC and rhodamine or separate filters. Cells that have bound Annexin V-FITC will show bright green staining on the plasma membrane. Cells which have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane).

TESTING RESULTS: Jurkat cells (treated with 2 µM camptothecin for 6 hours) were collected for annexin V assay according to the kit instructions. Results show 40-60 % apoptotic cells as analyzed by flow cytometry.
Restrictions For Research Use only
Format Liquid
Concentration 0.15 mg/mL
Buffer PBS containing 1 % BSA, 0.02 % sodium azide
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze!
Storage 4 °C
Expiry Date 12 months
Product cited in: Dondeti, Wubbenhorst, Lal et al.: "Integrative genomic analyses of sporadic clear cell renal cell carcinoma define disease subtypes and potential new therapeutic targets." in: Cancer research, Vol. 72, Issue 1, pp. 112-21, 2012 (PubMed).

Neri, Ren, Gratton et al.: "Bortezomib-induced "BRCAness" sensitizes multiple myeloma cells to PARP inhibitors." in: Blood, Vol. 118, Issue 24, pp. 6368-79, 2011 (PubMed).

Peters, Raghavan: "Endoplasmic reticulum calcium depletion impacts chaperone secretion, innate immunity, and phagocytic uptake of cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 187, Issue 2, pp. 919-31, 2011 (PubMed).

Sumitomo, Nakata, Higashino et al.: "Streptolysin S contributes to group A streptococcal translocation across an epithelial barrier." in: The Journal of biological chemistry, Vol. 286, Issue 4, pp. 2750-61, 2011 (PubMed).

Robbins, Tersey, Ogihara et al.: "Inhibition of deoxyhypusine synthase enhances islet {beta} cell function and survival in the setting of endoplasmic reticulum stress and type 2 diabetes." in: The Journal of biological chemistry, Vol. 285, Issue 51, pp. 39943-52, 2010 (PubMed).

Sharmeen, Skrtic, Sukhai et al.: "The antiparasitic agent ivermectin induces chloride-dependent membrane hyperpolarization and cell death in leukemia cells." in: Blood, Vol. 116, Issue 18, pp. 3593-603, 2010 (PubMed).

Wu, Zhou, Robertson et al.: "Identification of a bone marrow-derived CD8αα+ dendritic cell-like population in inflamed autoimmune target tissue with capability of inducing T cell apoptosis." in: Journal of leukocyte biology, Vol. 88, Issue 5, pp. 849-61, 2010 (PubMed).

Tsujii, Eguchi, Chenchik et al.: "Screening of cell death genes with a mammalian genome-wide RNAi library." in: Journal of biochemistry, Vol. 148, Issue 2, pp. 157-70, 2010 (PubMed).

Snyder, Reynolds, Hollingsworth et al.: "Clara cells attenuate the inflammatory response through regulation of macrophage behavior." in: American journal of respiratory cell and molecular biology, Vol. 42, Issue 2, pp. 161-71, 2010 (PubMed).

Radke, Reynolds, Melo et al.: "Mature human eosinophils express functional Notch ligands mediating eosinophil autocrine regulation." in: Blood, Vol. 113, Issue 13, pp. 3092-101, 2009 (PubMed).

Tu, Ren, Wang et al.: "The p53-cathepsin axis cooperates with ROS to activate programmed necrotic death upon DNA damage." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, Issue 4, pp. 1093-8, 2009 (PubMed).

Sumi, Ishida, Okumura et al.: "Activation of tumour-necrosis factor-related apoptosis-inducing ligand receptor enhances the severity of murine allergic conjunctivitis." in: The British journal of ophthalmology, Vol. 93, Issue 1, pp. 110-5, 2008 (PubMed).

Zhang, Wang, Liu et al.: "Regulation of tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by DJ-1 in thyroid cancer cells." in: Endocrine-related cancer, Vol. 15, Issue 2, pp. 535-44, 2008 (PubMed).

Vempati, Diaz, Barrientos et al.: "Role of cytochrome C in apoptosis: increased sensitivity to tumor necrosis factor alpha is associated with respiratory defects but not with lack of cytochrome C release." in: Molecular and cellular biology, Vol. 27, Issue 5, pp. 1771-83, 2007 (PubMed).

Terashima, Kojima, Fujimiya et al.: "The fusion of bone-marrow-derived proinsulin-expressing cells with nerve cells underlies diabetic neuropathy." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, Issue 35, pp. 12525-30, 2005 (PubMed).

Obeng, Boise: "Caspase-12 and caspase-4 are not required for caspase-dependent endoplasmic reticulum stress-induced apoptosis." in: The Journal of biological chemistry, Vol. 280, Issue 33, pp. 29578-87, 2005 (PubMed).

Okamoto, Kawamura, Kanbe et al.: "Suppression of serum IgE response and systemic anaphylaxis in a food allergy model by orally administered high-dose TGF-beta." in: International immunology, Vol. 17, Issue 6, pp. 705-12, 2005 (PubMed).

Tyurina, Serinkan, Tyurin et al.: "Lipid antioxidant, etoposide, inhibits phosphatidylserine externalization and macrophage clearance of apoptotic cells by preventing phosphatidylserine oxidation." in: The Journal of biological chemistry, Vol. 279, Issue 7, pp. 6056-64, 2004 (PubMed).

Oshiro, Landowski, Catlett-Falcone et al.: "Inhibition of JAK kinase activity enhances Fas-mediated apoptosis but reduces cytotoxic activity of topoisomerase II inhibitors in U266 myeloma cells." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 7, Issue 12, pp. 4262-71, 2001 (PubMed).

Grad, Bahlis, Reis et al.: "Ascorbic acid enhances arsenic trioxide-induced cytotoxicity in multiple myeloma cells." in: Blood, Vol. 98, Issue 3, pp. 805-13, 2001 (PubMed).

Zelphati, Wang, Kitada et al.: "Intracellular delivery of proteins with a new lipid-mediated delivery system." in: The Journal of biological chemistry, Vol. 276, Issue 37, pp. 35103-10, 2001 (PubMed).

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Catalog No. ABIN412007
1,122.00 $
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1000 tests
1,122.00 $

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