Annexin V Kit | Kits

Dot-plot showing Ramos cells staining with Annexin V:FITC (ANNEX100F) versus Propidium Iodide resulting in three distinct populations

Dot-plot showing Ramos cells staining with Annexin V:FITC (ANNEX100F) versus Propidium Iodide resulting in three distinct populations

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  • Dot-plot showing Ramos cells staining with Annexin V:FITC (ANNEX100F) versus Propidium Iodide resulting in three distinct populations
  • Annexin V:Biotin Assay Kit
  • Annexin V:FITC Assay Kit
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  • Product Type
    Kits
2 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    ANNEX100BF 100 Tests
    ANNEX100B
    ANNEX100FF 100 Tests
    ANNEX100F
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • This test employs the property of Annexin V:Biotin to bind to the membrane phospholipid phosphatidylserine (PS) in the presence of Ca2+. PS is exposed at the cell surface during the early stages of apoptosis. Detection of PS is a very sensitive method for detecting cells entering apoptosis, at a time point considerably ahead of nuclear changes such as DNA degradation.

      The conjugation protocol used to prepare Annexin V:Biotin has not changed the native phospholipid binding properties of Annexin V. This protocol is designed to measure apoptosis easily and quickly in a sample of suspended cells.
    • Preservative Stabilisers
      0.02%Sodium Azide (NaN3)
      1%Bovine Serum Albumin
      0.02%Sodium Azide (NaN3)
      1%Bovine Serum Albumin
    • Reagents In The Kit
      Annexin V:Biotin1 x 0.5ml vial
      Propidium Iodide1 x 1.6ml vial at 20ug/ml
      Binding buffer1 x 50ml vial at x 4 concentrate


      Note: This assay also requires streptavidin:FITC conjugate for visualisation (not supplied - see recommended useful reagents section).
      Annexin V:FITC1 x 0.5ml vial
      Propidium Iodide1 x 1.6ml vial at 20ug/ml
      Binding buffer1 x 50ml vial at x 4 concentrate
    • Storage
      Store at +4oC. DO NOT FREEZE.
      This product should be stored undiluted. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC. DO NOT FREEZE.
      This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      12 months from date of despatch.
      12 months from date of despatch.
    • GO Terms
      blood coagulation
      negative regulation of coagulation
      cytoplasm
      anti-apoptosis
      phospholipase inhibitor activity
      signal transduction
      calcium ion binding
      calcium-dependent phospholipid binding
    • UniProt
    • Entrez Gene
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNo
      Flow Cytometry
    • Application NameYesNo
      Flow Cytometry

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
      Please view the datasheet pdf for suggested dilutions.
    • Instructions For Use
      1) Dilute the binding buffer 1:4 in distilled water (50ml binding buffer + 150ml distilled water).

      2) Wash cells in PBS by gentle shaking or pipetting up and down.

      3) Resuspend cells in 200ul pre-diluted binding buffer, adjusting to a cell density of 2-5 x 105 cells/ml.

      4) Add 5ul Annexin V:Biotin to 195ul of the cell suspension prepared in step 3.

      5) Mix and incubate for 15 minutes at room temperature.

      6) Wash cells twice in 190ul of pre-diluted binding buffer.

      7) Resuspend cells in 190ul pre-diluted binding buffer.

      8) Add streptavidin:FITC conjugate.

      9) Mix and incubate for 30 minutes in the dark, at room temperature.

      10) Wash cells in 200ul pre-diluted binding buffer.

      11) Resuspend cells in 190ul pre-diluted binding buffer.

      12) Add 10ul of the Propidium Iodide solution.

      13) Analyse by flow cytometry.

      The flow cytometer is preferably set such that the Mean Fluorescence Intensity of the Annexin V negative population is between 1 and 10. Optimal parameter settings can be found using a positive control. For a positive control, incubate the cells with 3% formaldehyde in buffer during 30 minutes on ice. Wash away the formaldehyde and suspend the cells in cold binding buffer at 2-5 x 105 cells/ml. Proceed with step 2 as described above.
    • Instructions For Use
      1) Dilute the binding buffer 1:4 in distilled water (50ml binding buffer +150ml distilled water)

      2) Wash cells in PBS by gentle shaking or by pipetting up and down.

      3) Resuspend cells in 200ul pre-diluted binding buffer, adjusting to a cell density of 2-5 x 105 cells/ml.

      4) Add 5ul Annexin V:FITC to 195ul of the cell suspension prepared in step 3.

      5) Mix and incubate for 10 minutes in the dark, at room temperature.

      6) Wash cells in 200ul of pre-diluted binding buffer.

      7) Resuspend cells in 190ul pre-diluted binding buffer.

      8) Add 10ul of the Propidium Iodide solution.

      9) Analyse by flow cytometry.

      The flow cytometer is preferably set such that the Mean Fluorescence Intensity of the Annexin V negative population is between 1 and 10. Optimal parameter settings can be found using a positive control. For a positive control, incubate the cells with 3% formaldehyde in buffer during 30 minutes on ice. Wash away the formaldehyde and suspend the cells in cold binding buffer at 2-5 x 105 cells/ml. Proceed with step 2 as described above.

    Additional Annexin V Kit Formats

    FormatsApplicationsSizes available
    Annexin V Kit : BiotinF100 Tests
    Annexin V Kit : FITCF100 Tests
    • Copyright © 2014 Bio-Rad AbD Serotec

    Recommended Secondary Antibody

    Recommended Negative Isotype Control

    Useful Reagents

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Annexin V:Biotin Assay KitANNEX300B300 TestsF
      ANNEX300B
      Streptavidin:FITC7100021 mgF
      710002
      Streptavidin:FITCSTAR2B1 mgC, F, P
      STAR2B
      Annexin V:FITC Assay KitANNEX100F100 TestsF
      ANNEX100F
      Annexin V:FITC Assay KitANNEX300F300 TestsF
      ANNEX300F
      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Annexin V:Biotin Assay KitANNEX100B100 TestsF
      ANNEX100B
      Annexin V:Biotin Assay KitANNEX300B300 TestsF
      ANNEX300B
      Annexin V:FITC Assay KitANNEX300F300 TestsF
      ANNEX300F

      Recommended Positive Controls

      Histology Controls

        • Product Images

        • Dot-plot showing Ramos cells staining with Annexin V:FITC (ANNEX100F) versus Propidium Iodide resulting in three distinct populations
      • Application NameReference Images
        Flow Cytometry

      References

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