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Annexin V-FITC Apoptosis Kit

Details for Product No. ABIN411977, Supplier: Log in to see
Antigen
  • 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
Alternatives
Chemical Annexin V ELISA Kit
Reactivity
Chemical
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Application
Apoptosis Detection (AD), Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose Annexin V Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidylserine (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. The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
Sample Type Biological Fluids, Adherent Cell Culture, 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 Annexin V-FITC Apoptosis Detection Kit: Simple One Step Assay to Detect Apoptosis within 10 min. Can also differentiate Apoptosis vs Necrosis.
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. The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
Components Annexin V-FITC
1X Binding Buffer
Propidium Iodide (PI)
Alternative Name Annexin V (ANXA5 ELISA Kit Abstract)
Target Type Chemical
Background Annexin V Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane phosphatidylserine (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. The kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining.
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Application Notes Detect early/middle stages of apoptosis, differentiate apoptosis from necrosis.
Comment

Further details regarding sample type: Living cells (suspension and adherent)

Assay Time 0.5 h
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
Handling Advice Protect from light. Product is photosensitive.
Storage 4 °C
Expiry Date 12 months
Product cited in: Clarke, Weist, Walsh, Tenner: "Complement protein C1q bound to apoptotic cells suppresses human macrophage and dendritic cell-mediated Th17 and Th1 T cell subset proliferation." in: Journal of leukocyte biology, Vol. 97, Issue 1, pp. 147-60, 2015 (PubMed).

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Javeed, Sagar, Dutta, Smyrk, Lau, Bhattacharya, Truty, Petersen, Kaufman, Chari, Mukhopadhyay: "Pancreatic Cancer-Derived Exosomes Cause Paraneoplastic β-cell Dysfunction." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 21, Issue 7, pp. 1722-33, 2015 (PubMed).

Sasano, Mabuchi, Kuroda, Kawano, Matsumoto, Takahashi, Hisamatsu, Sawada, Hashimoto, Isobe, Testa, Kimura: "Preclinical Efficacy for AKT Targeting in Clear Cell Carcinoma of the Ovary." in: Molecular cancer research : MCR, Vol. 13, Issue 4, pp. 795-806, 2015 (PubMed).

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Liu, Fan, Tang, Ke: "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, Vol. 38, Issue 3, pp. 683-93, 2014 (PubMed).

Ahn, Coh, Lee, Shin, Kang, Youn: "Human adipose tissue-derived mesenchymal stem cells inhibit melanoma growth in vitro and in vivo." in: Anticancer research, Vol. 35, Issue 1, pp. 159-68, 2014 (PubMed).

Bergin, Reeves, Hurley, Wolfe, Jameel, Fitzgerald, McElvaney: "The circulating proteinase inhibitor α-1 antitrypsin regulates neutrophil degranulation and autoimmunity." in: Science translational medicine, Vol. 6, Issue 217, pp. 217ra1, 2014 (PubMed).

Cheng, Holloway, Nguyen, McCauley, Landesman, Kauffman, Shacham, Altura: "XPO1 (CRM1) inhibition represses STAT3 activation to drive a survivin-dependent oncogenic switch in triple-negative breast cancer." in: Molecular cancer therapeutics, Vol. 13, Issue 3, pp. 675-86, 2014 (PubMed).

Ganapathy, Xiao, Yang, Qi, Choi, Ha, Little, Yuan: "A low-dose arsenic-induced p53 protein-mediated metabolic mechanism of radiotherapy protection." in: The Journal of biological chemistry, Vol. 289, Issue 8, pp. 5340-7, 2014 (PubMed).

Ho, Tung, Kuo, Lin, Wu: "Ferruginol inhibits non-small cell lung cancer growth by inducing caspase-associated apoptosis." in: Integrative cancer therapies, Vol. 14, Issue 1, pp. 86-97, 2014 (PubMed).

Hurley, Lacey, ODwyer, Bergin, McElvaney, OBrien, McElvaney, Reeves, McElvaney: "Alpha-1 antitrypsin augmentation therapy corrects accelerated neutrophil apoptosis in deficient individuals." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 193, Issue 8, pp. 3978-91, 2014 (PubMed).

Islam, Thomas, Murty, Sedoris, Miller: "c-Myc quadruplex-forming sequence Pu-27 induces extensive damage in both telomeric and nontelomeric regions of DNA." in: The Journal of biological chemistry, Vol. 289, Issue 12, pp. 8521-31, 2014 (PubMed).

Ito, Horikawa, Suzuki, Kawauchi, Tanaka, Suzuki, Suzuki: "Human NAT10 is an ATP-dependent RNA acetyltransferase responsible for N4-acetylcytidine formation in 18 S ribosomal RNA (rRNA)." in: The Journal of biological chemistry, Vol. 289, Issue 52, pp. 35724-30, 2014 (PubMed).

Kim, Kim, Park, Park, Lee, Lee, Choe, Kim, Jeoung: "miR-326-histone deacetylase-3 feedback loop regulates the invasion and tumorigenic and angiogenic response to anti-cancer drugs." in: The Journal of biological chemistry, Vol. 289, Issue 40, pp. 28019-39, 2014 (PubMed).

Li, Duan, Jia, Bai, Zhang, Wang: "Flavonoids from tartary buckwheat induce G2/M cell cycle arrest and apoptosis in human hepatoma HepG2 cells." in: Acta biochimica et biophysica Sinica, Vol. 46, Issue 6, pp. 460-70, 2014 (PubMed).

Li, Cai, Xia, Yao, Chen, Zhang, Naranmandura, Liu, Wu: "Involvement of endoplasmic reticulum stress in all-trans-retinal-induced retinal pigment epithelium degeneration." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 143, Issue 1, pp. 196-208, 2014 (PubMed).

Muenyi, Trivedi, Helm, States: "Cisplatin plus sodium arsenite and hyperthermia induces pseudo-G1 associated apoptotic cell death in ovarian cancer cells." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 139, Issue 1, pp. 74-82, 2014 (PubMed).

Noguchi, Saito, Horie, Mikami, Suzuki, Morishita, Ohshima, Abiko, Mattsson, König, Lohr, Edlund, Botling, Micke, Nagase: "An integrative analysis of the tumorigenic role of TAZ in human non-small cell lung cancer." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 20, Issue 17, pp. 4660-72, 2014 (PubMed).