Annexin V-APC/7-AAD細胞凋亡檢測試劑盒

簡要描述：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.

產品型號：abs50008
廠商性質：生產廠家
更新時間：2025-06-13
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細胞生物學

重組蛋白原代細胞分離細胞培養細胞檢測細胞/組織處理外泌體研究其它動物血清 3D/類器官培養

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詳細介紹

品牌	absin	貨號	abs50008
規格	25T；100T	供貨周期	現貨
應用領域	化工,生物產業,制藥/生物制藥

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：

	25T	100T
Annexin V-APC	150ul	500ul
7-AAD	300ul	1ml
5×Binding buffer	5ml	15ml
Apoptosis Positive Control Solution	5ml	5ml

應用：

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(1×106-3×106cells), 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 5μL of Annexin V-APC, and the third one adds 5 μL 7-AAD 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.

B.Sample detection
1. Dilute 6 mL 5× binding buffer with 24 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 1×106cells/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 7-AAD 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.