CBA060 Annexin V-PE Apoptosis Detection Kit

CBA060
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Overview

Replacement Information

Key Specifications Table

Detection Methods
Fluorescence

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CBA060-100TEST
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      Plastic ampoule 100 test
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      A convenient kit useful for the identification of phosphatidylserine (PS) that is translocated from the cytoplasmic face of the plasma membrane to the cell surface during apoptosis. Annexin V has a strong natural affinity for PS, hence, it is used for detecting apoptosis. This assay can be performed on live cells.
      Catalogue NumberCBA060
      Brand Family Calbiochem®
      Materials Required but Not Delivered Apoptosis inducer
      Flow cytometer or fluorescence microscope
      References
      ReferencesDarzynkiewicz, Z., et al. 1997. Cytometry 27, 1.
      Frey, T. 1997. Cytometry 28, 253.
      Boersma, A.W.M., et al. 1996. Cytometry 24, 123.
      White, E. 1996. Genes and Devel. 10, 1.
      Homburg, C.H., et al. 1995. Blood 85, 532.
      Martin, S.J., et al. 1995. J. Exp. Med. 182, 1545.
      Vermes, I., et al. 1995. J. Immunol. Methods 184, 39.
      Koopman, G., et al. 1994. Blood 84 1415.
      Wyllie, A.H.,1993. Br. J. Cancer 67, 205.
      Darzynkiewicz, Z., et al. 1992. Cytometry 13, 795.
      Fadok, V.A., et al. 1992. J. Immunology 148, 2207.
      Kerr, J.F.R., et al. 1972. Cancer 26, 239.
      Product Information
      Detection methodFluorescence
      Form25 or 100 Tests
      FormatFlow cytometry or fluorescence microscopy
      Kit containsAnnexin V-PE, 1X Binding Buffer, and a user protocol.
      Quality LevelMQ100
      Applications
      Key Applications Flow Cytometry
      Immunofluorescence
      Biological Information
      Assay time15-30 min
      Sample TypeIntact cells
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Intended useThe Calbiochem® Annexin V-PE Apoptosis Detection Kit is a non-isotopic system for the detection of cell membrane alterations that accompany programmed cell death. Detection may be performed by flow cytometry or fluorescence microscopy.
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage +2°C to +8°C
      Storage ConditionsUpon arrival store the entire contents of the kit at 4°C.
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Kit containsAnnexin V-PE, 1X Binding Buffer, and a user protocol.
      Specifications

      Documentation

      Annexin V-PE Apoptosis Detection Kit Certificates of Analysis

      TitleLot Number
      CBA060

      References

      Reference overview
      Darzynkiewicz, Z., et al. 1997. Cytometry 27, 1.
      Frey, T. 1997. Cytometry 28, 253.
      Boersma, A.W.M., et al. 1996. Cytometry 24, 123.
      White, E. 1996. Genes and Devel. 10, 1.
      Homburg, C.H., et al. 1995. Blood 85, 532.
      Martin, S.J., et al. 1995. J. Exp. Med. 182, 1545.
      Vermes, I., et al. 1995. J. Immunol. Methods 184, 39.
      Koopman, G., et al. 1994. Blood 84 1415.
      Wyllie, A.H.,1993. Br. J. Cancer 67, 205.
      Darzynkiewicz, Z., et al. 1992. Cytometry 13, 795.
      Fadok, V.A., et al. 1992. J. Immunology 148, 2207.
      Kerr, J.F.R., et al. 1972. Cancer 26, 239.
      User Protocol

      Revision20-October-2008 RFH
      Form25 or 100 Tests
      FormatFlow cytometry or fluorescence microscopy
      Detection methodFluorescence
      StorageUpon arrival store the entire contents of the kit at 4°C.
      Intended useThe Calbiochem® Annexin V-PE Apoptosis Detection Kit is a non-isotopic system for the detection of cell membrane alterations that accompany programmed cell death. Detection may be performed by flow cytometry or fluorescence microscopy.
      BackgroundApoptosis is a programmed cell death that serves an important regulatory function during normal development, in homeostasis, and in some disease processes. In normal viable cells phosphatidylserine (PS) is located at the cytoplasmic surface of the cell membrane. Upon induction of apoptosis, rapid alterations in the organization of phospholipids in most cell types occur, leading to exposure of PS on the cell surface. Recognition of PS by phagocytes in vivo results in the removal of cells that are programmed to die, thus apoptosis is not commonly associated with the local inflammatory response that accompanies necrosis. In vitro detection of externalized PS can be achieved through interaction with the anticoagulant annexin V. In the presence of calcium, rapid high affinity binding of annexin V to PS occurs. PS translocation to the cell surface precedes nuclear breakdown, DNA fragmentation, and the appearance of most apoptosis-associated molecules making annexin V binding a marker of early-stage apoptosis.
      Principles of the assayThe Calbiochem® Annexin V-PE Apoptosis Detection Kit utilizes annexin V conjugated to phycoerythrin to bind to early apoptotic cells. Detection of bound annexin V is visualized by flow cytometry or fluorescence microscopy. The kit is supplied in two sizes, 100 tests or 25 tests.
      Materials providedThe 100 test kit contains the following components:

      • Annexin V-PE (Kit Component No. JA9205-500UL): 1 vial, 500 µl
      • 1X Binding Buffer (Kit Component No. JA9206-50ML): 1 bottle, 50 ml

      The 25 test kit contains the following components:

      • Annexin V-PE (Kit Component No. JA9207-125UL): 1 vial, 125 µl
      • 1X Binding Buffer (Kit Component No. JA9208-12.5ML): 1 bottle, 12.5 ml
      Materials Required but not provided Apoptosis inducer
      Flow cytometer or fluorescence microscope
      Precautions and recommendations When using adherent cells for flow cytometry analysis gently trypsinize and wash the cells once with serum-containing cell culture medium prior to incubation with Annexin V-PE.
      When using adherent cells for fluorescence microscopy, grow the cells directly on a coverslip; following incubation with Annexin V-PE invert the coverslip on a glass slide for visualization. The cells can also be washed and fixed in 2% formaldehyde prior to analysis; however, cells must first be incubated with Annexin V-PE prior to fixation as any cell membrane disruption can cause nonspecific binding of Annexin V to PS on the inner surface of the cell membrane.
      Cells that are stained with Annexin V-PE will appear orange-red around the plasma membrane.
      Detailed protocol1. Induce apoptosis using a desired method.
      2. Count the cells and collect 1-5 x 105 cells by centrifugation.
      3. Resuspend the cells with 500 µl 1X Binding Buffer.
      4. Add 5 µl Annexin V-PE and incubate at room temperature for 5 min in the dark.
      5. Analyze the samples by flow cytometry or fluorescence microscopy.
      a. Flow cytometry: analyze annexin V-PE by flow cytometry (excitation max: 488 nm; emission max: 578 nm) using the phycoerythrin emission signal detector (usually FL2).
      b. Fluorescence microscopy: place cell suspension on a glass slide and cover with a cover slip. For adherent cells, invert the cover slip on a glass slide; analyze the samples using a fluorescence microscope equipped with a rhodamine filter.
      Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
      Interactive Pathways™ is a trademark of EMD Chemicals, Inc.