Annexin V/PI double staining was performed using an Annexin-V-FLUOS Staining Kit (Roche Applied Science, 11988549001) according to the manufacturer’s protocol. .. The cells were analyzed by flow cytometry (FACScan, Becton Dickinson) and the data were evaluated using Cell Quest software.

Protocol 1: Annexin V staining preliminary preparations. Prepare binding buffer ( BB): 140 mM NaCl, 4 mM KCl, 0.75 mM MgCl2 and 10 mM HEPES in DDW.

2. Bones cleaned and crushed in ice cold HBSS + 2% FBS ina motar and pestle. 3. Filter cells through 40m filter to remove bone fragments into 50 mL Annexin V Binding Buffer Prepare 1X Binding Buffer by diluting the Cell-Based Assay Annexin V Binding Buffer (10X) (Item No. 600302) 1:10 in distilled water. Mix well and keep at room temperature. The diluted 1X Binding Buffer will be stable for one year at room temperature. Annexin V FITC/Propidium Iodide Staining Solution Annexin-V measurements Direct fluorescence staining of apoptotic cells for flow cytometric analysis was performed with the Annexin-V-FLUOS staining Kit (Roche).

Annexin v staining

After the indicated times, 5 × 105 –1 × 106 cells were harvested by trypsin, washed twice with PBS, stained with both Annexin-V-FLUOS and propidium iodide and analysed in a flow cytometer. 2016-07-09 Annexin V/PI double staining was performed using an Annexin-V-FLUOS Staining Kit (Roche Applied Science, 11988549001) according to the manufacturer’s protocol. .. The cells were analyzed by flow cytometry (FACScan, Becton Dickinson) and the data were evaluated using Cell Quest software.

○. Studies on 2 Sep 2019 Annexin V is a 35 kDa phospholipid-binding protein. Its strong calcium- dependent affinity for phosphatidylserine (PS) can be used to identify These cells will stain with Annexin V but not with viability dyes, thus distinguishing cells in early apoptosis.

因此AnnexinV被作为检测细胞早期凋亡的灵敏指标之一。. 碘化丙啶（Propidium Iodide,PI）是一种核酸染料，它不能透过完整的细胞膜，但凋亡中晚期的细胞和死细胞由于细胞膜通透性的增加，PI能够透过细胞膜而使细胞核染红。. 因此将Annexin V与PI匹配使用，就可以将处于不同凋亡时期的细胞区分开来。. 因此，将Annexin V与PI联合使用时，PI 则被排除在活细胞（Annexin V-/PI-）和早期

2. Transfer 100 µL of cell suspension in 5 mL test tube. 3. Add 5 µL of PE Annexin V. 4.

Superfolder GFP :: Fluorescent Protein Database Foto. Characterization of Annexin V Fusion with the Superfolder Foto. Gå till. Figure 5 | Soluble expression of

Analysis is by flow cytometry or fluorescence microscopy.

Add annexin V and calcium only 10 minutes before acquisition.
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Annexin V staining protocol for apoptosis A. Incubation of cells with annexin V-FITC. Induce apoptosis by the desired method. Collect 1–5 x 10 5 cells by B. Quantification by flow cytometry. Analyze annexin V-FITC binding by flow cytometry (Ex = 488 nm; Em = 350 nm) using C. Detection by Apoptotic cells have a minimal uptake of PI and will appear dimly stained.

Use only binding buffer (BB). After taking the cells from the culture keep them on ice at all times. Prepare a stock solution of annexin V and PI in BB, so that every sample will receive 10 μL from the stock.
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Protocol 2: Annexin V staining experimental protocol Use only binding buffer (BB). After taking the cells from the culture keep them on ice at all times. Prepare a stock solution of annexin V and PI in BB, so that every sample will receive 10 μL from the stock.

The process of PS translocation occurs prior to the loss of membrane integrity. Company Telephone: Fax: Hours: Monday to Friday 8:30 - 17:30 PST (GMT-8) Location: 520 Mercury Drive Annexin V-FITC and PI Staining Solution should be stored in the dark. Cautions 1. For maximal assay performance, this kit should be used within 12 months.