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
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 FITC
Application
Apoptosis Induction (AI), Flow Cytometry (FACS), Fluorescence Microscopy (FM), Light Microscopy (LM)
Pubmed 21 references available
Quantity 100 tests
Options
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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
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
Research Area Cardiovascular, Atherosclerosis, Apoptosis/Necrosis, Coagulation
Application Notes Detect early/middle stages of apoptosis, differentiate apoptosis from necrosis.
Comment

Flow cytometry (Ex = 488 nm, Em = 530 nm) and fluorescence microscopy
Simple one step staining procedure in 10 minutes
Fast and convenient
Kit can differentiate apoptosis vs necrosis when performing both Annexin V-FITC and PI staining

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
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Yanamala, Kapralov, Djukic et al.: "Structural re-arrangement and peroxidase activation of cytochrome c by anionic analogues of vitamin E, tocopherol succinate and tocopherol phosphate." in: The Journal of biological chemistry, Vol. 289, Issue 47, pp. 32488-98, 2014 (PubMed).

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Yang, Wang, Qin et al.: "JKA97, a novel benzylidene analog of harmine, exerts anti-cancer effects by inducing G1 arrest, apoptosis, and p53-independent up-regulation of p21." in: PLoS ONE, Vol. 7, Issue 4, pp. e34303, 2012 (PubMed).

Xiong, Hong, Du et al.: "Roles of STAT3 and ZEB1 proteins in E-cadherin down-regulation and human colorectal cancer epithelial-mesenchymal transition." in: The Journal of biological chemistry, Vol. 287, Issue 8, pp. 5819-32, 2012 (PubMed).

Liu, Vaithiyalingam, Shi et al.: "BID binds to replication protein A and stimulates ATR function following replicative stress." in: Molecular and cellular biology, Vol. 31, Issue 21, pp. 4298-309, 2011 (PubMed).

Greenlee-Wacker, Briseño, Galvan et al.: "Membrane-associated CD93 regulates leukocyte migration and C1q-hemolytic activity during murine peritonitis." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 187, Issue 6, pp. 3353-61, 2011 (PubMed).

Xie, Kondeti, Lin et al.: "Role of extracellular matrix renal tubulo-interstitial nephritis antigen (TINag) in cell survival utilizing integrin (alpha)vbeta3/focal adhesion kinase (FAK)/phosphatidylinositol 3-kinase ..." in: The Journal of biological chemistry, Vol. 286, Issue 39, pp. 34131-46, 2011 (PubMed).

Wu, Hour, Leung et al.: "2-(Naphthalene-1-yl)-6-pyrrolidinyl-4-quinazolinone inhibits skin cancer M21 cell proliferation through aberrant expression of microtubules and the cell cycle." in: The Journal of pharmacology and experimental therapeutics, Vol. 338, Issue 3, pp. 942-51, 2011 (PubMed).

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Catalog No. ABIN411977
396.00 $
Plus shipping costs $45.00
Quantity
Price
100 tests
396.00 $

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