Annexin V-Cy5 Apoptosis Kit

Antigen: Annexin A5 (ANXA5)

Synonyme: PP4, ANX5, ENX2, Anx5, R74653, LC5, MGC93655, ANXA5, MGC126971, anx, anx5, ANX V, anxa5, cb989, wu:fa98f06, wu:fj10f10, MGC89158

Konjugat: Cy5

Applikation: Durchflusszytometrie (FACS), Fluoreszenzmikroskopie (FM)

Produktnummer: ABIN411983

Produktbeschreibung

Weitere Bezeichnung: Annexin V

Beschreibung: 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.

Anwendungen

Protokoll: A. Incubation of Cells with Annexin V-Cy5: 1. Induce apoptosis by desired methods. 2. Collect 1-5 x 10 5 cells by centrifugation. 3. Resuspend cells in 500 μ l of 1X Annexin V Binding Buffer. 4. Add 5 μ l 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 cells by flow cytometry (Ex = 649 nm, Em = 670 nm) using a Helium-Neon Laser. For adherent cells, trypsinize and gently wash cells with serum-containing medium 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, 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% formaldehyde before visualization. (Cells must be incubated with Annexin V-Cy5 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 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. IV. Related Products: Apoptosis Detection Kits & Reagents Annexin V Kits & Bulk Reagents Caspase Assay Kits & Reagents Mitochondrial Apoptosis Kits & Reagents Nuclear Apoptosis Kits & Reagents Apoptosis Inducers and Set Apoptosis siRNA Vectors Cell Fractionation System Mitochondria/Cytosol Fractionation Kit Nuclear/Cytosol Fractionation Kit Membrane Protein Extraction Kit Cytosol/Particulate Rapid Separation Kit Mammalian Cell Extraction Kit FractionPREP Fractionation System Cell Proliferation & Senescence Quick Cell Proliferation Assay Kit Senescence Detection Kit High Throughput Apoptosis/Cell Viability Assay Kits LDH-Cytotoxicity Assay Kit Bioluminescence Cytotoxicity Assay Kit Live/Dead Cell Staining Kit Cell Damage & Repair HDAC & HAT Fluorometric & Colorimetric Assays & Drug Discovery Kits DNA Damage Quantification Kit Glutathione & Nitric Oxide Fluorometric & Colorimetric Assay Kits Signal Transduction cAMP & cGMP Assay Kits Akt & JNK Activity Assay Kits Beta-Secretase Activity Assay Kit Adipocyte & Lipid Transfer Recombinant Adiponectin, Survivin, & Leptin CETP & PLTP Activity Assay & Drug Discovery Kits Total Cholesterol Quantification Kit Molecular Biology & Reporter Assays siRNA Vectors Cloning Insert Quick Screening Kit Mitochondrial & Genomic DNA Isolation Kits 5 Minutes DNA Ligation Kit 20 Minutes Gel Staining/Destaining Kit β -Galactosidase Staining Kit & Luciferase Reporter Assay Kit Growth Factors and Cytokines Adiponectin/Resistin/Leptin and their Antibodies Recombinant Protein A and Protein G Recombinant Complement C5a Recombinant Cytokines and Growth Factors Monoclonal and Polyclonal Antibodies Components K103-25 K103-100 K103-400 25 assays 100 assays 400 assays Annexin V-Cy5 125 μ l 500 μ l 2 ml 1X Binding Buffer 12.5 ml 50 ml 2 x 100 ml

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: 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).