产品介绍
1. 外观：黄色粉末
2. 纯度：≥99% (HPLC) 超纯级
3. 产品参数：Ex(nm) 352 Em(nm) 461 Solvent: Water
产品描述
Hoechst 33258是可透过细胞膜并对DNA染色的细胞核染色试剂，在嵌入双链DNA后释放强烈的蓝色荧光。Hoechst 33258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。Hoechst 33258-DNA的激发和发射波长分别为352 nm和461 nm。
使用方法
1、对于固定的细胞或组织：
1) 对于细胞或组织样品，固定后，适当洗涤去除固定剂。随后如果需要进行免疫荧光染色，则先进行免疫荧光染色，染色完毕后再按后续步骤进行Hoechst 33258染色。如果不需要进行其它染色，则直接进行后续的Hoechst 33258染色。
2) 对于贴壁细胞或组织切片，加入少量Hoechst 33258染色液，覆盖住样品即可。对于悬浮细胞，至少加入待染色样品体积3倍的染色液，并混匀。室温放置3-5分钟。
3) 吸除Hoechst 33258染色液，用TBST 、PBS或生理盐水洗涤2-3次，每次3-5分钟。
4) 直接在荧光显微镜下或封片后荧光显微镜下观察。观察细胞凋亡时，会看到凋亡细胞的细胞核呈致密浓染，或呈碎块状致密浓染。
2、对于活细胞或组织：
5) 加入适当量Hoechst 33258染色液，必须充分覆盖住待染色的样品，通常对于六孔板一个孔需加入1ml染色液，对于96 孔板一个孔需加入100微升染色液。
6) 在适宜于细胞培养的温度培养20-30分钟。弃染色液，用PBS或培养液洗涤2-3次即可进行荧光检测。
储存条件：-20℃避光保存，有效期一年。
注意事项
1)Hoechst 33258对人体有一定刺激性，请注意适当防护。
2)荧光染料都存在淬灭的问题，建议染色后尽量当天完成检测。
3)为减缓荧光淬灭可以使用抗荧光淬灭封片液。
4)为了您的安全和健康，请穿实验服并戴一次性手套操作。
Hoechst 33258 is a nuclear staining reagent that can penetrate cell membranes and stain DNA. It releases strong blue fluorescence after intercalating double-stranded DNA. Hoechst 33258 is often used for cell apoptosis detection. After staining, observe with a fluorescence microscope or detect with a flow cytometer. The excitation and emission wavelengths of Hoechst 33258-DNA are 352 nm and 461 nm, respectively.
Instructions
1. For fixed cells or tissues
1) For cell or tissue samples, after fixation, wash them appropriately to remove fixative. Then, if immunofluorescence staining is required, perform immunofluorescence staining first, and then follow the subsequent steps for Hoechst 33258 staining after the staining is completed. If no other staining is required, the subsequent Hoechst 33258 staining is directly performed.
2) For adherent cells or tissue sections, add a small amount of Hoechst 33258 staining solution to cover the sample. For suspended cells, add at least 3 times the volume of the sample to be stained with staining solution and mix well. Leave at room temperature for 3-5 minutes.
3) Aspirate the Hoechst 33258 staining solution, wash 2-3 times with TBST, PBS or saline, 3-5 minutes each time.
4) Observe directly under a fluorescence microscope or after mounting the slide. When observing cell apoptosis, the nucleus of apoptotic cells will be densely stained, or fragmented and densely stained.
2. For living cells or tissues
5) Add an appropriate amount of Hoechst 33258 staining solution, which must fully cover the sample to be stained. Usually, 1ml staining solution is needed for one well of a six-well plate, and 100 microliters of staining solution is required for one well of a 96-well plate.
6) Incubate for 20-30 minutes at a temperature suitable for cell culture. Discard the staining solution, wash 2-3 times with PBS or culture solution to perform fluorescence detection.
Storage conditions: Store at -20°C and protected from light, valid for one year.
Precautions
1) Hoechst 33258 is irritating to the human body, please pay attention to proper protection.
2) Fluorescent dyes have the problem of quenching. It is recommended to complete the test on the same day after dyeing.
3) To slow down fluorescence quenching, anti-fluorescence quenching mounting solution can be used.
4) For your safety and health, please wear lab coats and disposable gloves for operation.
G2410289
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