Annexin V-APC/PI apoptosis analysis kit

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产品描述：

产品名称：Annexin V-APC/PI apoptosis analysis kit

储存/保存方法: Keep as concentrated solution. Store at 4°C and protected from prolonged exposure to light. Do not freeze.The validity period is 6 months.

描述:

Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Fluorochrome-labeled Annexin V can then be used to specifically target and identify apoptotic cells. Annexin V Binding Buffer is recommended for use with Annexin V staining.

Products List：Annexin V-APC、Propidium Iodide solution、Annexin V Binding Buffer、Apoptosis Positive Control Solution

应用: Each lot of this reagent is quality control tested by immunofluorescent staining with flow cytometric analysis (The amount of the reagent is suggested to be used 5 μl/10^5 cells). Since applications vary, the appropriate dilutions must be determined for individual use.

使用方法: Suggested Staining Protocol A.Parameters regulation 1. Harvest cell(1×10⁶ -3×10⁶ cells), then sepatate the cells in two parts. Wash cells with cold PBS, then centrifuge the cells and disgard the supernatant. 2. Suspend one part of cells in 200μL 1× binding buffer, store at 4℃ for use. 3. Suspend the other part of cells in 500μL Apoptosis Positive Control Solution, and incubate for 10 minutes in room temperature. Wash cells with more than 3.0 mL cold PBS, blot the supernatant, then suspend the cells in 200μL 1×binding buffer. 4. Mix the two parts cells together, then separate the cells in three tubes, and add 100 μL of cells in each tube. 5. The first tube is Blank Control, the second one adds 5 μL of Annexin V-APC, and the third one adds 5 μL PI solution. 6. Gently vortex each tube and incubate for 5 minutes in room temperature, protected from light. 7. Before analyzing by flow cytometry, using the blank control and single dye sample to regulate voltage and compensation, as shown in Figure Parameters regulation. B.Sample detection 1. Dilute 3 mL 10×binding buffer with 27 mL distilled water for 10 tests. 2. Harvest cell（about 1×10⁶cells per test）then wash with cold PBS. 3. Suspend cells in 1 mL 1× Binding Buffer, 300×g centrifugation for 10 minutes, then remove the Binding Buffer from the cell pellet. 4. Resuspend cells in 1 mL 1× Binding Buffer , adjust cell concentration to 1×10⁶6cells/mL. 5. Add 100 μL of cells (1×10⁵cells) to each labeled tube. 6. Add 5 μL of Annexin V-APC to appropriate tubes. 7. Gently vortex each tube and incubate for 10 minutes in room temperature, protected from light. 8. Add 5 μL PI solution incubation for 5min in room temperature, protected from light. 9. Add PBS to 500μL and vortex gently. 10. Analyze by flow cytometry in 1 hour.

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