Annexin V-EGFP Reagent

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      Catalog number:E-CK-A119

      Size:
      • 50T
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      Price: $110

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      Introduction:

      Elabscience® Annexin V-EGFP is developed to identify apoptotic and necrotic cells.

      Annexin V is a member of the annexin family, which binds to phosphatidylserine (PS) in a calcium-dependent manner.  The fluorescent format of this intracellular protein, Annexin V-EGFP, binds specifically to the PS on outer leaflet of cell membrane by flow cytometry or fluorescence microscopy.

      Cells at different apoptotic stages can be distinguished by using Annexin V and DNA dyes like Propidium Iodide (PI) or 7-AAD.



      Components:


      Cat.

      Products

      100 T

      200 T

      500T

      1000 T

      Storage

       E-CK-A119

      Annexin V-EGFP

      500 μL

      1 mL

      2.5 mL

      5 mL

      2-8

                                       Manual

       One Copy





      Jurkat cells were treated with 1μM Camptothecin and detected by this reagent and PI:






      Jurkat cells were treated with 1μM Camptothecin (Left) or not (Right) for 4 h. Annexin V-EGFP single-positive cells were early apoptotic cells, Annexin V-EGFP and PI double-positive cells were necrotic or late apoptotic cells, and PI single-positive cells were nude nuclear cells.




      Staining Procedure:

      1.  Induce apoptosis of suspension cells with reagents of interest. Collect cell cultures and centrifuge at 300 g for 5 min, discard the supernatant. Add appropriate PBS to wash the cells, resuspend gently and count the cells.

      Note: This product is only validated in suspension cells. Good cell viability is the key to the experiment. When the adherent cells are used for apoptotic detection, treatments like digestion may increase the ratio of necrotic or apoptotic cells and cause uncontrollable effects on the experimental results. Please be aware!

      2. Split the cell suspension into tubes, 1-5 × 105 cells for each. Centrifuge at 300 g for 5 min, discard the supernatant. Add appropriate PBS to wash the cells and discard the supernatant. Add 500 μL of 1 × Annexin V Binding Buffer [#Cat:E-CK-A151] to resuspend the cells.

      3.  Add 5 µl of Annexin V-EGFP and 5 µl of DNA dye (PI [#Cat:E-CK-A161] or 7-AAD[#Cat:E-CK-A162]) to each tube.

      4. Gently vortex the cells and incubate at room temperature for 15-20 min in the dark.

      5. Analyze the cells immediately with proper machine settings. Otherwise, place the cells on ice in the dark

      and analyze within 1 h.


      Storage:

      Store at 4°C for up to one year in dark.


      Cautions:

      1.  For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.

      2.  Detect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis.

      3.  Avoid extended exposure of the samples to direct light to protect the fluorophores from quenching.

      4.  For your safety and health, please wear the lab coat and disposable gloves before the experiments.




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