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Annexin V-FITC Apoptosis Kit

Details for Product No. ABIN411977
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Target Name (Antigen)
Synonyms anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158, ANX5, ENX2, PP4, RPRGL3, Anx5, R74653, LC5, enx2
Reactivity
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(2), (5), (5), (16), (2), (2), (7), (4), (7), (4), (1), (4), (6), (2)
Application
Flow Cytometry (FACS), Fluorescence Microscopy (FM)
Pubmed 21 references available
Quantity 100 tests
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Catalog No. ABIN411977
368.50 $
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Purpose Detects early/middle stages of apoptosis, differentiates apoptosis from necrosis
Sample Type Cell Samples
Detection Method Fluorometric
Specificity The assay is based on the observation that soon after initiating apoptosis, most cell types translocate the membrane phospholipid phosphatidylserine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can easily be detected by staining with a fluorescent conjugate of Annexin V, a protein that has a strong natural affinity for PS.
Characteristics The one-step staining procedure takes only 10 minutes. Detection can be analyzed by flow cytometry or by fluorescence microscopy. The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
Components Annexin V-FITC (500 µL)
1X Binding Buffer (50 mL)
Alternative Name Annexin V
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 method.
2. Collect 1-5 x 10^5 cells by centrifugation.
3. Resuspend cells in 500 µL of 1X Binding Buffer.
4. 4. Add 5 µL of Annexin V-FITC and 5 µL of propidium iodide (PI 50 µg/mL, optional).
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-FITC 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 analyzing adherent cells, gently trypsinize and wash cells once with serum-containing media 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. 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.
Note: Cells must be incubated with Annexin V-FITC 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 that have bound Annexin V-FITC will show green staining in the plasma membrane. Cells that 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).
Restrictions For Research Use only
Storage 4 °C
Expiry Date 12 months
Product cited in: Gao, Liu, Du et al.: "Implication of oxygen-regulated protein 150 (ORP150) in apoptosis induced by proteasome inhibitors in human thyroid cancer cells." in: The Journal of clinical endocrinology and metabolism, Vol. 95, Issue 11, pp. E319-26, 2010 (PubMed).

Liu, Li, Raisch: "Clotrimazole induces a late G1 cell cycle arrest and sensitizes glioblastoma cells to radiation in vitro." in: Anti-cancer drugs, Vol. 21, Issue 9, pp. 841-9, 2010 (PubMed).

Yamaguchi, Kobayashi, Narita et al.: "Sonodynamic therapy using water-dispersed TiO(2)-polyethylene glycol compound on glioma cells: Comparison of cytotoxic mechanism with photodynamic therapy." in: Ultrasonics sonochemistry, Vol. 18, Issue 5, pp. 1197-204, 2011 (PubMed).

Harashima, Inao, Imamura et al.: "Roles of the PI3K/Akt pathway and autophagy in TLR3 signaling-induced apoptosis and growth arrest of human prostate cancer cells." in: Cancer immunology, immunotherapy : CII, 2011 (PubMed).

Naderi, Moazzeni, Pourfathollah et al.: "High expression of fas ligand on cord blood dendritic cells: a possible immunoregulatory mechanism after cord blood transplantation." in: Transplantation proceedings, Vol. 43, Issue 10, pp. 3913-9, 2011 (PubMed).

Austgen, Johnson, Park et al.: "The adaptor protein CRK is a pro-apoptotic transducer of endoplasmic reticulum stress." in: Nature cell biology, Vol. 14, Issue 1, pp. 87-92, 2011 (PubMed).

Fransen, Berden, Koeter et al.: "Effect of administration of apoptotic blebs on disease development in lupus mice." in: Autoimmunity, 2012 (PubMed).

Xia, Dai, Li et al.: "The protective effect of sonic hedgehog is mediated by the propidium iodide 3-kinase/AKT/BCL-2 pathway in cultured rat astrocytes under oxidative stress." in: Neuroscience, 2012 (PubMed).

Shi, Qian, Li et al.: "Nuclear Translocation of β-catenin is Essential for Glioma Cell Survival." in: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, 2012 (PubMed).

Sela, Molotski, Golan et al.: "Human Embryonic Stem Cells Exhibit Increased Propensity to Differentiate During the G1 Phase Prior to Phosphorylation of pRB." in: Stem cells (Dayton, Ohio), 2012 (PubMed).

Liu, Fan, Tang et al.: "The Resveratrol Attenuates Ethanol-Induced Hepatocyte Apoptosis Via Inhibiting ER-Related Caspase-12 Activation and PDE Activity In Vitro." in: Alcoholism, clinical and experimental research, 2013 (PubMed).

General Cao, Szabolcs, Dutta et al.: "Neuropilin-1 mediates divergent R-Smad signaling and the myofibroblast phenotype." in: The Journal of biological chemistry, Vol. 285, Issue 41, pp. 31840-8, 2010 (PubMed).

Chung, Moon, Jung et al.: "Potentiation of Bortezomib-Induced Apoptosis by TGF-{beta} in Cultured Human Tenon's Fibroblasts: Contribution of the PI3K/Akt Signaling Pathway." in: Investigative ophthalmology & visual science, Vol. 51, Issue 12, pp. 6232-7, 2010 (PubMed).

Awasthi, Schwarz, Schwarz: "Enhancing cytotoxic agent activity in experimental pancreatic cancer through EMAP II combination therapy." in: Cancer chemotherapy and pharmacology, 2010 (PubMed).

Liu, Bertram, Shi et al.: "Proapoptotic Bid mediates the Atr-directed DNA damage response to replicative stress." in: Cell death and differentiation, 2010 (PubMed).

Li, Allen, Bonanno: "SOLUBLE ADENYLYL CYCLASE MEDIATES BICARBONATE -DEPENDENT CORNEAL ENDOTHELIAL CELL PROTECTION." in: American journal of physiology. Cell physiology, 2010 (PubMed).

Niu, Wang, Li et al.: "Cord blood-derived cytokine-induced killer cells biotherapy combined with second-line chemotherapy in the treatment of advanced solid malignancies." in: International immunopharmacology, 2011 (PubMed).

Awasthi, Kirane, Schwarz et al.: "Smac mimetic-derived augmentation of chemotherapeutic response in experimental pancreatic cancer." in: BMC cancer, Vol. 11, Issue 1, pp. 15, 2011 (PubMed).

Thambiayya, Wasserloos, Huang et al.: "LPS-induced decrease in intracellular labile zinc [Zni] contributes to apoptosis in cultured sheep pulmonary artery endothelial cells (SPAEC)." in: American journal of physiology. Lung cellular and molecular physiology, 2011 (PubMed).

Hunsucker, Magarotto, Kuhn et al.: "Blockade of interleukin-6 signalling with siltuximab enhances melphalan cytotoxicity in preclinical models of multiple myeloma." in: British journal of haematology, 2011 (PubMed).

Otsuka, Mizuki, Fujita et al.: "Constitutively Active FGFR3 with Lys650Glu Mutation Enhances Bortezomib Sensitivity in Plasma Cell Malignancy." in: Anticancer research, Vol. 31, Issue 1, pp. 113-22, 2011 (PubMed).

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