DAPI dihydrochloride(4',6-联脒-2-苯基吲哚二盐酸盐)是一种可与富含A-T序列的DNA结合的荧光染料。
别名:4',6-Diamidino-2-phenylindole dihydrochloride
DAPI是一种可对DNA 染色的细胞核染色试剂,它在嵌入双链DNA后释放蓝色荧光。DAPI常用于细胞凋亡检测,染色后用荧光显微镜观察或流式细胞仪检测。尽管DAPI不能通过活细胞膜,但却能穿透扰乱的细胞膜而对核染色。DAPI具有很高的光漂白承受水平,能用来检测酵母线粒体DNA,叶绿体DNA,病毒DNA,microplasm DNA以及染色体DNA。DAPI-DNA复合物的激发和发射波长分别为360nm和460nm。
Protocols for Counterstaining with DAPI:
Immunofluorescence Staining
Note: Cells may be fixed by method of choice, such as with 4% paraformaldehyde. DAPI staining is done after staining for other markers. Fixation and permeabilization of cells is not necessary to counterstain with DAPI.
1. Fix cells using method of choice.
2. Incubate the cells with phosphate buffered saline (PBS) for 15 minutes.
3. Dilute the DAPI solution (we recommend to 300 nM) with PBS. Remove excess PBS from the slide and cover with DAPI solution, making sure the cells are completely covered.
4. Incubate for up to 5 minutes. Rinse the slide several times to remove all free DAPI. It is recommended to use a mounting medium with antifade reagent to reduce fluorescence quenching. The slide is ready to view under the microscope with appropriate filters.
Nuclear Staining for Flow Cytometry
Note: Cells may be fixed with 4% paraformadehyde or absolute ethanol.
1. Fix cells with absolute ethanol or 4% paraformaldehyde.
2. Centrifuge, discard the supernatant, and add 5 ml of PBS, allowing cells to rehydrate for 15 minutes.
3. Dilute the DAPI to 3 µM (recommended) in staining buffer.
4. Centrifuge cell suspension from step 3 above, discard supernatant, and add 1 ml of DAPI solution, then incubate at room temperature for 15 minutes.
5. Analyze by flow cytometry using an instrument with appropriate lasers.
注意事项:
1.DAPI对人体有一定刺激性,请注意适当防护。Adv Mater. 2023 Feb 22;e2211138.
Genetically Programmable Vesicles for Enhancing CAR-T Therapy Against Solid Tumors
Nucleic Acids Res. 2022 Aug 12;50(14):7938-7958.
Oncol Lett. 2022 May;23(5):163.
Appl Biochem Biotechnol Appl Biochem Biotechnol. 2022 Jul 1.
J Nanobiotechnology. 2021 Nov 25;19(1):391.
Ann Biomed Eng. 2021 Jun;49(6):1551-1560.
Theranostics. 2020 Apr 27;10(13):5671-5686.
Aging (Albany NY). 2020 Jun 9;12(11):11025-11041.
Life Sci. 2020 Sep 15;257:118021.
分子量 | 350.25 |
分子式 | C16H17Cl2N5 |
CAS号 | 28718-90-3 |
溶解性(25°C) | Water 5 mg/mL (Need ultrasonic) |
储存条件 | -20°C, protect from light, sealed |
运输方式 | 冰袋运输,根据产品的不同,可能会有相应调整。 |
*下述溶液配置方法仅为基于分子量计算出的理论值。不同产品在配置溶液前,需考虑其在不同溶剂中的溶解度限制。
Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
---|---|---|---|
1 mM | 2.8551 mL | 14.2755 mL | 28.551 mL |
5 mM | 0.571 mL | 2.8551 mL | 5.7102 mL |
10 mM | 0.2855 mL | 1.4276 mL | 2.8551 mL |
小鼠 | 大鼠 | 兔 | 豚鼠 | 仓鼠 | 狗 | |
重量 (kg) | 0.02 | 0.15 | 1.8 | 0.4 | 0.08 | 10 |
体表面积 (m2) | 0.007 | 0.025 | 0.15 | 0.05 | 0.02 | 0.5 |
Km 系数 | 3 | 6 | 12 | 8 | 5 | 20 |
动物 A (mg/kg) = 动物 B (mg/kg) × | 动物 B的Km系数 |
动物 A的Km系数 |
例如,依据体表面积折算法,将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量,需要将20 mg/kg 乘以小鼠的Km系数(3),再除以大鼠的Km系数(6),得到化合物用于大鼠的等效剂量为10 mg/kg。
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