Annexin V-Cy5 Apoptosis Kit

Details for Product No. ABIN411983
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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
Chemical
(34), (5), (4), (3), (16), (2), (3), (15), (7), (5), (10), (4), (1), (5), (10), (2)
Conjugate
This Annexin V antibody is conjugated to Cy5
Application
Apoptosis Induction (AI), Flow Cytometry (FACS), Fluorescence Microscopy (FM), Light Microscopy (LM)
Pubmed 19 references available
Quantity 100 tests
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Purpose Annexin V Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidyl-serine (PS) from the inner face of the plasma membrane to the cell surface. Once on the cell surface, PS can be easily detected by staining with a fluorescent conjugate of Annexin V, a protein that has a high affinity for PS. The one-step staining procedure takes only 10 minutes. Detection can be analyzed by flow cytometry or by fluorescence microscopy.
Sample Type Biological Fluids
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 Annexin V-Cy5 Apoptosis Detection Kit: Simple One Step Assay to Detect Apoptosis within 10 min.
Detection: Flow Cytometry or Fluorescence Microscopy
The one-step staining procedure takes only 10 minutes. Detection can be analyzed by flow cytometry or by fluorescence microscopy.
Components Annexin V-Cy5
1X Binding Buffer
Alternative Name Annexin V (ANXA5 ELISA Kit Abstract)
Target Type Chemical
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Application Notes Optimal working dilution should be determined by the investigator.
Comment

Flow cytometry (Ex = 649 nm, Em = 670 nm) and fluorescence microscopy
Simple one step staining procedure in 10 minutes
Fast and convenient
Annexin V-Cy5 will show bright red-blue staining on the plasma membrane

Assay Time 0.5 h
Protocol A. Incubation of cells with Annexin V-Cy5
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. Add 5 µL 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 Annexin V-Cy5 binding by flow cytometry (Ex = 649 nm, Em = 670 nm) using a Helium-Neon Laser.
For analyzing adherent cells, gently trypsinize and wash cells once with serum-containing media 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. 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. (Cells must be incubated with Annexin V-Cy3 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 Cy5 filter, or FITC/Cy3/Cy5 triple band filter (Chroma Technology) if performing triple labeling using these dyes, or detect cells using a CCD camera. Cells that have bound Annexin V-Cy5 will show bright red-blue staining on the plasma membrane.
Restrictions For Research Use only
Handling Advice Protect from light. Product is photosensitive.
Storage 4 °C
Expiry Date 12 months
Product cited in: Samejima, Ogawa, Ageichik et al.: "Auxin-induced rapid degradation of inhibitor of caspase-activated DNase (ICAD) induces apoptotic DNA fragmentation, caspase activation, and cell death: a cell suicide module." in: The Journal of biological chemistry, Vol. 289, Issue 45, pp. 31617-23, 2014 (PubMed).

Yamaguchi, Maruyama, Urade et al.: "Immunosuppression via adenosine receptor activation by adenosine monophosphate released from apoptotic cells." in: eLife, Vol. 3, pp. e02172, 2014 (PubMed).

Dyugovskaya, Berger, Polyakov et al.: "The development of giant phagocytes in long-term neutrophil cultures." in: Journal of leukocyte biology, Vol. 96, Issue 4, pp. 511-21, 2014 (PubMed).

Gulshan, Brubaker, Wang et al.: "Sphingomyelin depletion impairs anionic phospholipid inward translocation and induces cholesterol efflux." in: The Journal of biological chemistry, Vol. 288, Issue 52, pp. 37166-79, 2013 (PubMed).

Kontos, Kourtis, Dane et al.: "Engineering antigens for in situ erythrocyte binding induces T-cell deletion." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 1, pp. E60-8, 2013 (PubMed).

Segawa, Suzuki, Nagata: "Constitutive exposure of phosphatidylserine on viable cells." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, Issue 48, pp. 19246-51, 2011 (PubMed).

Weidinger, Krause, Mueller et al.: "FOXO3 is inhibited by oncogenic PI3K/Akt signaling but can be reactivated by the NSAID sulindac sulfide." in: The Journal of clinical endocrinology and metabolism, Vol. 96, Issue 9, pp. E1361-71, 2011 (PubMed).

Klagge, Weidinger, Krause et al.: "The role of FOXO3 in DNA damage response in thyrocytes." in: Endocrine-related cancer, Vol. 18, Issue 5, pp. 555-64, 2011 (PubMed).

Wang, Martin, Harris et al.: "Caspase inhibition blocks cell death and enhances mitophagy but fails to promote T-cell lymphoma." in: PLoS ONE, Vol. 6, Issue 5, pp. e19786, 2011 (PubMed).

Du, Wolf, Schafer et al.: "BH3 domains other than Bim and Bid can directly activate Bax/Bak." in: The Journal of biological chemistry, Vol. 286, Issue 1, pp. 491-501, 2011 (PubMed).

Martinez-Garcia, Popovic, Min et al.: "The MMSET histone methyl transferase switches global histone methylation and alters gene expression in t(4;14) multiple myeloma cells." in: Blood, Vol. 117, Issue 1, pp. 211-20, 2011 (PubMed).

Aldaz, Escudero, Freeman: "Live imaging of Drosophila imaginal disc development." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, Issue 32, pp. 14217-22, 2010 (PubMed).

Huang, Yamada, Kidoya et al.: "EphB4 overexpression in B16 melanoma cells affects arterial-venous patterning in tumor angiogenesis." in: Cancer research, Vol. 67, Issue 20, pp. 9800-8, 2007 (PubMed).

Zhang, Lawson, Polosukhin et al.: "Inhibitor of differentiation 1 promotes endothelial survival in a bleomycin model of lung injury in mice." in: The American journal of pathology, Vol. 171, Issue 4, pp. 1113-26, 2007 (PubMed).

Collingwood, Smirnova, Bogush et al.: "T-cell ubiquitin ligand affects cell death through a functional interaction with apoptosis-inducing factor, a key factor of caspase-independent apoptosis." in: The Journal of biological chemistry, Vol. 282, Issue 42, pp. 30920-8, 2007 (PubMed).

Brown, Martin, Maurice et al.: "Caspase inhibition blocks cell death and results in cell cycle arrest in cytokine-deprived hematopoietic cells." in: The Journal of biological chemistry, Vol. 282, Issue 4, pp. 2144-55, 2007 (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).

Nagy, Barcza, Gonchoroff et al.: "Nitric oxide-dependent mitochondrial biogenesis generates Ca2+ signaling profile of lupus T cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 173, Issue 6, pp. 3676-83, 2004 (PubMed).

Breckenridge, Nguyen, Kuppig et al.: "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).

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Catalog No. ABIN411983
396.00 $
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Quantity
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100 tests
396.00 $

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