耗材描述
BODIPY FL C5-Ceramide
产品活性:高尔基体是由单位膜构成的扁平囊叠加在一起所组成。扁平囊为圆形,边缘膨大且具有穿孔。 高尔基体荧光探针是一种 BODIPY 标记的神经酰胺衍生物。神经酰胺的合成发生在内质网,高尔基体荧光探针可以通过神经酰胺转运蛋白(CERT)或囊泡转运输送到高尔基体,从而实现染料的特异性标记。BODIPY Fl C5-Ceramide 是一种高尔基体特异性绿色荧光染料,可实现单个细胞可视化。Ex/Em=505 nm/512 nm。
产品来源: https://www.medchemexpress.cn/bodipy-fl-c5-ceramide.html
研究领域:Others
作用靶点:Fluorescent Dye
In Vitro: General Protocol
1 Preparation of Golgi working solution
1.1 Preparation of the stock solution
Dissolve Golgi in DMSO to obtain 5 mM of Golgi.
Note: It is recommended to store the stock solution at -20°C -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Golgi working solution
Dilute the stock solution in HBSS to obtain 1-10 μM of Golgi working solution.
Note: Please adjust the concentration of Golgi working solution according to the actual situation.
2 Cell staining
2.1 Suspension cells (6-well plate)
a.Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL.
b.Add 1 mL of working solution, and then incubate at room temperature for 20-30 minutes.
c.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d.Wash twice with PBS, 5 minutes each time.
e.Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a.Culture adherent cells on sterile coverslips.
b.Remove the coverslip from the medium and aspirate excess medium.
c.Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 20-30 minutes.
d.Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.
Storage
-20℃ 1 year.Protect from light.
Precautions
1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.
2. Please adjust the concentration of Golgi 1-43 working solution according to the actual situation.
3. when the cell culture solution is removed with poor results, the cells can be washed with an appropriate amount of Hanks balanced salt solution.
4. This product is for R&D use only, not for drug, household, or other uses.
5. For your safety and health, please wear a lab coat and disposable gloves to operate.
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