pSIVA™ Flow Cytometry Kit

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  • pSIVA™ REAL-TIME Apoptosis Flow Cytometry Kit
  • pSIVA™ REAL-TIME Apoptosis Flow Cytometry Kit
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  • Product Type
    Kits
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    APO003BFdatasheet pdfdatasheet pdf datasheet pdf datasheet pdf datasheet pdf100 Tests
    APO003B
    APO003AFdatasheet pdfdatasheet pdf datasheet pdf datasheet pdf datasheet pdf25 Tests
    APO003A
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • The process of apoptosis is undertaken in several stages defined by specific cellular morphologies. One of the earlier stages of apoptosis is a change of the plasma membrane’s phospholipid asymmetry. This rearrangement results in the translocation of phosphatidylserine (PS) from the inner to the outer plasma membrane (in non-apoptotic cells PS is exclusively located to the inner plasma membrane). However, apoptosis is reversible until reaching a certain point in the pathway and until then PS exposure can be considered as a transient event. The event defining whether the cell can be rescued and continues living is the onset of mitochondrial outer membrane permeabilization (MOMP) (Chipuk et al. 2006). Prior to reaching this point, PS exposure may be transient as molecules can relocate back to the inner plasma membrane (a phenomenon known as “PS flipping”) (van der Mark et al. 2013).

      The pSIVA™ (polarity-Sensitive Indicator of Viability & Apoptosis) probe is a biosensor conjugated to the green emitting IANBD dye (excitation maximum 488 nm, emission maximum 530 nm) and only fluoresces when bound to PS in the presence of Ca2+ (Kim et al. 2010a, 2010b). The method thereby allows the analysis of kinetic apoptosis events by flow cytometry.
    • Intended Use
    • Product Form
    • Reconstitution
    • Preparation
    • Preservative Stabilisers
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
      Pack Size: 25 TestspSIVA™-IANBD 125 μl
      Propidium iodide staining solution 125 μl
      10X PBS buffer 5.0 ml
      10X Binding buffer 2.0 ml
      Pack Size: 100 TestspSIVA™-IANBD 500 μl
      Propidium iodide staining solution 500 μl
      10X PBS buffer 20.0 ml
      10X Binding buffer 5.0 ml
      Pack Size: 25 TestspSIVA™-IANBD 125 μl
      Propidium iodide staining solution 125 μl
      10X PBS buffer 5.0 ml
      10X Binding buffer 2.0 ml
      Pack Size: 100 TestspSIVA™-IANBD 500 μl
      Propidium iodide staining solution 500 μl
      10X PBS buffer 20.0 ml
      10X Binding buffer 5.0 ml
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
    • Storage
      Store at +4oC. DO NOT FREEZE.
      This product should be stored undiluted. This product is photosensitive and should be protected from light.
      Store at +4oC. DO NOT FREEZE.
      This product should be stored undiluted. This product is photosensitive and should be protected from light.
    • Shelf Life
      6 months from date of despatch
      6 months from date of despatch
    • Acknowledgements
      pSIVA is a trademark of Novus Biologicals and is protected under patent no. 8,541,549
      pSIVA is a trademark of Novus Biologicals and is protected under patent no. 8,541,549
    • 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 NameYesNoMin DilutionMax Dilution
      Flow Cytometry
      Refer to Instuctions For UsePack Size: 25 Tests
      Refer to Instructions For UsePack Size: 100 Tests
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry
      Refer to Instuctions For UsePack Size: 25 Tests
      Refer to Instructions For UsePack Size: 100 Tests

    • 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.
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • ELISA
    • ELISA
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
    • Instructions For Use
      1. Prepare 1X PBS solution by diluting 10X PBS concentrate 1/10 in distilled water and place on ice. Prepare a total of 2 ml 1X PBS per sample.

      2. Prepare 1X binding buffer solution by diluting 10X binding buffer concentrate 1/10 in distilled water. Prepare a total of 0.5 ml 1X binding buffer per sample.

      3. Centrifuge samples at 300 x g for 5 min. Discard the supernatant ensuring that the pellet is not disturbed.

      4. Wash pellet twice in 1.0 ml of cold 1X PBS. After each wash, centrifuge tubes at 300 x g for 5 min and discard the supernatant ensuring that the pellet is not disturbed.

      5. Using 1X binding buffer, resuspend the pellet to a concentration 1 x 106 to 1 x 107 cells/ml. Resuspend the pellet carefully by flicking the sample tube, DO NOT VORTEX.

      6. For one color pSIVA-IANBD staining, add 5 μl pSIVA-IANBD to 100 μl of the cell suspension. For two color staining, add both 5 μl pSIVA-IANBD + 5 μl PI to 100 μl of the cell suspension. Carefully mix by swirling the sample tube.

      7. Ensure sample tubes are protected from light and incubate for 20 min at room temperature (approximately 20-25°C).

      8. Add 400 μl of 1X binding buffer to each sample tube.

      9. Analyze cells by flow cytometry immediately using the FL1 channel for pSIVA-IANBD (excitation maximum 488 nm, emission maximum 530 nm) and the FL2 channel for PI (excitation maximum 535 nm, emission maximum 617 nm).

      * The stated pSIVA-IANBD and PI quantities are guidelines only and may have to be optimized.

      Instructions for use can be found at www.bio-rad-antibodies.com/static/uploads/ifu/apo003a.pdf
    • Instructions For Use
      1. Prepare 1X PBS solution by diluting 10X PBS concentrate 1/10 in distilled water and place on ice. Prepare a total of 2 ml 1X PBS per sample.

      2. Prepare 1X binding buffer solution by diluting 10X binding buffer concentrate 1/10 in distilled water. Prepare a total of 0.5 ml 1X binding buffer per sample.

      3. Centrifuge samples at 300 x g for 5 min. Discard the supernatant ensuring that the pellet is not disturbed.

      4. Wash pellet twice in 1.0 ml of cold 1X PBS. After each wash, centrifuge tubes at 300 x g for 5 min and discard the supernatant ensuring that the pellet is not disturbed.

      5. Using 1X binding buffer, resuspend the pellet to a concentration 1 x 106 to 1 x 107 cells/ml. Resuspend the pellet carefully by flicking the sample tube, DO NOT VORTEX.

      6. For one color pSIVA-IANBD staining, add 5 μl pSIVA-IANBD to 100 μl of the cell suspension. For two color staining, add both 5 μl pSIVA-IANBD + 5 μl PI to 100 μl of the cell suspension. Carefully mix by swirling the sample tube.

      7. Ensure sample tubes are protected from light and incubate for 20 min at room temperature (approximately 20-25°C).

      8. Add 400 μl of 1X binding buffer to each sample tube.

      9. Analyze cells by flow cytometry immediately using the FL1 channel for pSIVA-IANBD (excitation maximum 488 nm, emission maximum 530 nm) and the FL2 channel for PI (excitation maximum 535 nm, emission maximum 617 nm).

      * The stated pSIVA-IANBD and PI quantities are guidelines only and may have to be optimized.

      Instructions for use can be found at www.bio-rad-antibodies.com/static/uploads/ifu/apo003b.pdf

    Additional pSIVA™ Flow Cytometry Kit Formats

    Formats Applications Sizes available
    pSIVA™ Flow Cytometry Kit : IANBD (Green Fluorescence) F 100 Tests | 25 Tests
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          pSIVA™ REAL-TIME Apoptosis Fluorescent Microscopy KitAPO0041 KitIC, IF, LI
          APO004
          pSIVA™ REAL-TIME Apoptosis Flow Cytometry KitAPO003B100 TestsF
          APO003B
          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          pSIVA™ REAL-TIME Apoptosis Fluorescent Microscopy KitAPO0041 KitIC, IF, LI
          APO004
          pSIVA™ REAL-TIME Apoptosis Flow Cytometry KitAPO003A25 TestsF
          APO003A

          Recommended Positive Controls

            Histology Controls

              References

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