Annexin A5 (ANXA5) Protein

Details for Product No. ABIN412008
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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
Reactivity
(49), (15), (9), (4), (4), (4), (4), (3), (1), (1)
Host
(67), (10), (9), (2), (2)
Application
Flow Cytometry (FACS), Fluorescence Microscopy (FM)
Pubmed 21 references available
Quantity 200 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: Crișan, Cleophas, Oosting et al.: "Soluble uric acid primes TLR-induced proinflammatory cytokine production by human primary cells via inhibition of IL-1Ra." in: Annals of the rheumatic diseases, 2015 (PubMed).

Cleophas, Crişan, Lemmers et al.: "Suppression of monosodium urate crystal-induced cytokine production by butyrate is mediated by the inhibition of class I histone deacetylases." in: Annals of the rheumatic diseases, 2015 (PubMed).

Shetty, Matalia, Nuijts et al.: "Safety profile of accelerated corneal cross-linking versus conventional cross-linking: a comparative study on ex vivo-cultured limbal epithelial cells." in: The British journal of ophthalmology, Vol. 99, Issue 2, pp. 272-80, 2015 (PubMed).

Gao, Dong, Lin et al.: "Reduction of AUF1-mediated follistatin mRNA decay during glucose starvation protects cells from apoptosis." in: Nucleic acids research, Vol. 42, Issue 16, pp. 10720-30, 2014 (PubMed).

Herrmann, Cherryholmes, Schroeder et al.: "TLR9 is critical for glioma stem cell maintenance and targeting." in: Cancer research, Vol. 74, Issue 18, pp. 5218-28, 2014 (PubMed).

Gaudette, Iwakoshi, Boise: "Bcl-xL protein protects from C/EBP homologous protein (CHOP)-dependent apoptosis during plasma cell differentiation." in: The Journal of biological chemistry, Vol. 289, Issue 34, pp. 23629-40, 2014 (PubMed).

Murray, Gavile, Nair et al.: "CD28-mediated pro-survival signaling induces chemotherapeutic resistance in multiple myeloma." in: Blood, Vol. 123, Issue 24, pp. 3770-9, 2014 (PubMed).

Brentnall, Weir, Rongvaux et al.: "Procaspase-3 regulates fibronectin secretion and influences adhesion, migration and survival independently of catalytic function." in: Journal of cell science, Vol. 127, Issue Pt 10, pp. 2217-26, 2014 (PubMed).

Pal, Cao, Gaisina et al.: "Inhibition of GSK-3 induces differentiation and impaired glucose metabolism in renal cancer." in: Molecular cancer therapeutics, Vol. 13, Issue 2, pp. 285-96, 2014 (PubMed).

Liu, Shoji-Kawata, Sumpter et al.: "Autosis is a Na+,K+-ATPase-regulated form of cell death triggered by autophagy-inducing peptides, starvation, and hypoxia-ischemia." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 51, pp. 20364-71, 2013 (PubMed).

Tanaka-Nakanishi, Yasunaga, Takai et al.: "HTLV-1 bZIP factor suppresses apoptosis by attenuating the function of FoxO3a and altering its localization." in: Cancer research, Vol. 74, Issue 1, pp. 188-200, 2014 (PubMed).

Kim, Park, Kim et al.: "miR-200b and cancer/testis antigen CAGE form a feedback loop to regulate the invasion and tumorigenic and angiogenic responses of a cancer cell line to microtubule-targeting drugs." in: The Journal of biological chemistry, Vol. 288, Issue 51, pp. 36502-18, 2013 (PubMed).

Ori, Kato, Sanjo et al.: "Essential roles of K63-linked polyubiquitin-binding proteins TAB2 and TAB3 in B cell activation via MAPKs." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 190, Issue 8, pp. 4037-45, 2013 (PubMed).

Duan, Xu, Dong et al.: "Ectopic expression of miR-34a enhances radiosensitivity of non-small cell lung cancer cells, partly by suppressing the LyGDI signaling pathway." in: Journal of radiation research, Vol. 54, Issue 4, pp. 611-9, 2013 (PubMed).

Lindner, Concannon, Boukes et al.: "Systems analysis of BCL2 protein family interactions establishes a model to predict responses to chemotherapy." in: Cancer research, Vol. 73, Issue 2, pp. 519-28, 2013 (PubMed).

Zhang, Du, Meng et al.: "Beclin 1 enhances proteasome inhibition-mediated cytotoxicity of thyroid cancer cells in macroautophagy-independent manner." in: The Journal of clinical endocrinology and metabolism, Vol. 98, Issue 2, pp. E217-26, 2013 (PubMed).

Chen, Huang, Fan-Chiang et al.: "Apoptosis of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae), which failed to migrate within its natural host." in: The Journal of experimental biology, Vol. 216, Issue Pt 2, pp. 230-5, 2012 (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).

Bahlis, McCafferty-Grad, Jordan-McMurry et al.: "Feasibility and correlates of arsenic trioxide combined with ascorbic acid-mediated depletion of intracellular glutathione for the treatment of relapsed/refractory multiple myeloma." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 8, Issue 12, pp. 3658-68, 2002 (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. ABIN412008
385.00 $
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Quantity
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200 tests
385.00 $

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