产品说明书
FAQ
COA
已发表文献
Hoechst 33342 是一种可以穿透细胞膜的蓝色荧光染料，它在嵌入双链 DNA 后释放强烈的蓝色荧光，对细胞的毒性较低。Hoechst 33342 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。也可以用于普通的细胞核染色，或常规的DNA染色。
本品为粉末状，用于细胞核染色时，推荐的Hoechst 33342工作浓度为0.5-10 μg/mL。如需特定浓度或者经浓度优化的产品，可选购Hoechst 33342染液(1mg/ml)（Cat No.40732ES03）、Hoechst 33342染液(即用型)（Cat No. 40732ES10或Cat No. 40732ES50）。
产品性质
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中文名称（Chinese Synonym） |
二苯甲亚胺, 三氯化氢 2'-(4-乙基苯基)-5-(4-甲基-1-哌嗪基)-2,5'-二-1H-苯并咪唑 |
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英文名称（English Synonym） |
2’-(4-Ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5’-bi-1H-benzimidazole, trihydrochloride |
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分子式（Molecular Formula） |
C27H28N6O·3HCl |
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分子量（Molecular Weight） |
561.93 |
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CAS号（CAS NO.） |
23491-52-3 |
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荧光光谱（Fluorescence Spectral） |
Hoechst 33342的Ex/Em=346/460; Hoechst33342-核酸的Ex/Em=350/461. 荧光可用氙气灯，泵弧灯或者UV激光激发；可用DAPI滤光器、蓝色GFP滤光器、Alexa Fluor 350滤光器等检测。 |
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纯度（Purity） |
≥95%（HPLC） |
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结构（Structure） |
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运输与保存方法
冰袋（wet ice）运输。-20℃干燥避光保存，有效期2年。
注意事项
1）Hoechst 33342 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。
5）本产品仅作科研用途！
配制母液
Hoechst 33342溶于水，溶解度可达20 mg/mL。因此可用无菌超纯水配制成1-5 mg/mL的储存液于4℃避光保存。使用时用PBS或者其他合适缓冲液将其稀释成0.5-10 μg/mL工作液。
【注】：Hoechst 33342溶于PBS或者其他缓冲液时会有沉淀产生，因此不能用上述缓冲液进行储存液的配制。
使用方法
1. 用PBS或合适的缓冲液制备 0.5-10 μg/mL Hoechst 33342染色液。
2. 固定的细胞或组织染色：
对于固定的细胞或组织样品，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 33342染色。如果不需要进行其它染色，则直接进行后续的Hoechst 33342染色。
a）对于贴壁细胞或组织切片：加入适量Hoechst 33342染色液，覆盖住样品即可。
b）对于悬浮细胞：至少加入待测染色样品体积3倍的染色液，混匀。室温放置3-5分钟。
c）吸除Hoechst 33342染色液，用TBST、PBS或生理盐水洗涤2-3次，每次3-5分钟。
d）直接在荧光显微镜下观察或封片后荧光显微镜下观察。激发波长350 nm，发射波长460 nm。
3. 活细胞或组织染色：
a）细胞培养物中加入适量Hoechst 33342染色液，约1/10细胞培养基体积，必须充分覆盖待染色的样品。通常对于六孔板，每孔需加入1 mL染色液，对于96孔板，每孔需加入100 μL染色液。
b）在 37℃培养细胞 10～20 分钟。
c）用 PBS或合适的缓冲液洗细胞两次。
d）直接在荧光显微镜下观察或封片后荧光显微镜下观察。激发波长350 nm，发射波长460 nm。
HB230112
Q:如何做到同时染色溶酶体和细胞核的？
A:建议用 Hoechst 33258/ Hoechst 33342 染色细胞核，这个染料可以直接染活细胞不需要固定操作。
Q：DAPI和Hoechst染料相当类似，如何二选一？
A：DAPI是非常通用的蓝色荧光染料，用于细胞核复染。对固定和透化的细胞/组织，具有非常明亮的细胞核标记。然而，此染料被认为是半渗透至不渗透的染料，用于活细胞染色的结合不一致。Hoechst 33342是一种细胞膜渗透性的染料，具有同DAPI的结合机制和荧光特征。非常适合用于活细胞成像，效果就如同DAPI用于固定细胞染色。
Q：使用无血清培养基染色效果不好，荧光值很低，染色时间很久才能染上。
A：无血清培养基效果不是很好，建议使用PBS缓冲液染色，效果比较好。
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