耗材描述
Diphenyl Blue
产品活性:Diphenyl Blue (Trypan Blue) 是一种细胞活性染料,最常用的死细胞鉴定染料,常用于检测细胞膜的完整性和细胞的存活率。Diphenyl Blue 染色是组织和细胞培养中方法之一。细胞丧失活性或细胞膜不完整时,Diphenyl Blue 可将其染成蓝色,正常的活细胞细胞膜结构完整,能够排斥 Diphenyl Blue,细胞不会被染成蓝色。但巨噬细胞能够吞噬 Diphenyl Blue,所以可用于巨噬细胞的活体染色剂。
产品来源: https://www.medchemexpress.cn/Diphenyl_Blue.html
研究领域:Others
作用靶点:Fluorescent Dye
In Vitro: Preparation of Trypan Blue working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of Trypan Blue in 100 mL 0.85% Nacl to obtain 0.4% Trypan Blue.
Note: It is recommended to store the stock solution at -20℃ -80℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Trypan Blue working solution
Dilute the stock solution in 0.85% Nacl to obtain 0.04% of Trypan Blue working solution.
Note: Please adjust the concentration of Trypan Blue working solution according to the actual situation.
Cell staining
2.1 For suspension cells, centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells, discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of rypan Blue working solution, and then incubate at room temperature for 5 minutes.
2.3 Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope.
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