DAPI dilactate  (Synonyms: 4',6-Diamidino-2-phenylindole dilactate)

DAPI (dilactate) is a blue fluorescent dye that preferentially binds dsDNA and binds to minor groove AT clusters. DAPI (dilactate) is combined with dsDNA, and the fluorescence was enhanced about 20-fold. DAPI (dilactate) can be used to identify the cell cycle and specifically stains the nucleus but not the cytoplasm. DAPI (dilactate) form is more soluble in water than DAPI (dihydrochloride) form. ^[1][2]

一般方案
1. DAPI dilactate 工作液的制备
1.1 储备液的制备
将 5 mg DAPI dilactate 溶解于 1 mL ddH₂O 以获得 5 mg/mL 的储备液 (DAPI, dilactate 10.9 mM)。
注意：建议将储备液储存在 -20°C 或 -80°C 避光条件下，避免反复冻融。
1.2 DAPI dilactate 工作液的制备
将原液在PBS中以1:5000稀释至1 μg/mL。将工作溶液保存在4°C。
注意：请根据需要调整 DAPI dilactate 工作液的浓度
2、细胞染色
2.1 悬浮细胞 (96 孔板)
a．在 4°C 下以 1000 g 离心 3-5 分钟，然后弃去上清液。PBS洗涤2次，每次5分钟。细胞密度为1×10⁶/mL。
b．加入1 mL工作液，室温孵育3-10分钟。
c．4℃、400 g 离心 3-4 分钟，弃上清。
d．用 PBS 洗涤 2 次，每次 5 分钟。
e．用无血清细胞培养基或 PBS 重悬细胞。荧光显微镜或流式细胞术观察。
2.2 贴壁细胞
a．在无菌盖玻片上培养贴壁细胞。
b.从培养基上取下盖玻片并吸出多余的培养基。
c.加入100 μL工作液，轻摇使之完全覆盖细胞，室温孵育 3-10 分钟。
d．用培养基洗涤2次，每次 5 分钟。荧光显微镜或流式细胞仪观察。
注意事项
长期储存时，可将储备溶液等分并储存于 ≤-20°C。短期保存时，可将溶液置于 2-6°C 避光保存。如果处理得当，DAPI 溶液至少可稳定保存 6 个月。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

457.48

C₂₂H₂₇N₅O₆

28718-91-4

固体

Light yellow to yellow

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

• [1]. Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.  [Content Brief]

  [2]. Nazaroff CD, et al. Assessment of Lung Eosinophils In Situ Using Immunohistological Staining. Methods Mol Biol. 2021;2223:237-266.  [Content Brief]