DAPI是一种能够特异性结合到双链DNA（dsDNA）小沟的AT碱基对处，发出蓝色荧光的核酸染料。DAPI与DNA结合后，通过和小沟处的水分子置换，产生比自身强20多倍的荧光。DAPI也可以结合RNA，但结合的方式不同。相较于DAPI与dsDNA结合后的最大发射波长（~460 nm），DAPI/RNA具有较长的最大发射波长（~500 nm），其荧光亮度也仅有DAPI/dsDNA的20%。
本产品浓度为1.0 mg/mL，用于细胞核染色时，推荐工作浓度为0.1-10 μg/mL
注意事项：
1. DAPI具有致畸性，请注意适当防护。使用完的液体请合理处置丢弃。
2. 荧光染料存在淬灭的问题，建议染色后尽量当天完成检测。
使用说明：
1. 根据DAPI染色液应用的不同，建议使用前进行滴定以确定最佳工作浓度。
2. 用流式缓冲液稀释母液，配制成所需要的工作液（0.1-10 μg/mL）。
3. 活细胞染色：
a. 收集单细胞悬液，96孔板按照每孔100 μL 加入DAPI工作液；6孔板按照每孔1 mL加入DAPI工作液，使染色液充分覆盖样品；
b. 室温避光孵育10-15 min；
c. 直接进行流式分析或荧光显微镜分析。
4. 固定细胞染色：
a. 固定后，适当洗涤去除固定剂。在其它染色完成后，再进行DAPI染色;
b. 96孔板按照每孔加入100 μL DAPI工作液；6孔板按照每孔加入1 mL工作液，使染色液充分覆盖样品；
c. 室温避光孵育10-15 min；
d. 直接进行流式分析或荧光显微镜分析。
The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding of DAPI to dsDNA produces a ~20-fold fluorescence enhancement, apparently due to the displacement of water molecules from both DAPI and the minor groove. DAPI also binds RNA, however in a different binding mode. The DAPI/RNA complex exhibits a longer-wavelength fluorescence emission maximum than the DAPI/dsDNA complex (~500 nm versus ~460 nm) and a quantum yield that is only about 20% as high.
Attention:
1. DAPI is a known mutagen and should be handled with care. The dye must be disposed of safely and in accordance with applicable local regulations.
2. The fluorescent dye has the problem of quenching, and it is recommended to finish the analysis on the same day after dyeing.
Instructions for Use：
1. It is recommended to titrate the DAPI Staining Solution to obtain optimal results as applications vary.
2. Dilute the D1372407 with FACs buffer to the appropriate working solution (0.1-10 μg/mL).
3. Staining of Live Cells for Viability Analysis:
a. Obtain a single cell suspension, add 100 μL working solution to a 96-well and 1 mL to a 6-well;
b. Incubate 5 minutes at room temperature;
c. Proceed to analysis by flow cytometry or fluorescence analysis.
4. Staining of Fixed Cells for DNA Content Analysis:
a. Wash cells once to remove the fixation. DAPI staining is normally performed after all other staining;
b. Add appropriate volume of DAPI working solution (100 μL for a 96-well and 1 mL for a 6-well);
c. Incubate 5 minutes at room temperature;
d. Proceed to analysis by flow cytometry or fluorescence analysis.
ZJ25F1129308
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