Hoechst 33258
Cat.No：B8030 Solarbio
Appearance：Yellow or Green Powder
Storage：Store at -20℃，3 years
CAS：23491-45-4
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Endocrinology & Hormones
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HIF
Immunology & Inflammation
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Cell Cycle
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DNA Damage & DNA Repai
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Ubiquitin
JAK/STAT
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Oscillating Mixing & Stirring & Dispersing Instrument
Temperature Control & Sampling & Grinding Crushing Instrument
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Spectrophotometer
Centrifuge
Shaker
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Product Information
| Name | Hoechst 33258 |
| CAS | 23491-45-4 |
| Molecular Formula | C25H24N6O·3HCl |
| Molecular Weight | 533.88 |
| Purity/Grade | ≥98%(TLC) |
| Appearance | Yellow or Green Powder |
| Storage | Store at -20℃，3 years |
| Unit | Piece |
| Specification | 25mg |
别名：2'-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)- 2,5'-bi(1H-benzimidazole) trihydrochloride;2-[2- (4-Hydroxyphenyl)-6-benzimidazoyl]-6-(1-methyl-4- piperazyl)benzimidazole trihydrochloride;HOE 33258； Hoechst 33258
说明：荧光色素；用于DNA、染色体和核酸的染色；荧光染色法和免疫荧光技术。Hoechst 33258 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 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，也可以用于普通的细胞核染色，或常规的DNA染色。
物理性状：黄色至深黄色粉末；溶于水，黄或棕色溶液，参考浓度为9.80 - 10.20 mg/ml；熔点＞300℃。
用途：细胞遗传学研究的常用试剂，荧光染色细胞的DNA ，插入DNA的A-T区，具有低的细胞毒性，形成膜渗透性荧光DNA链。
用于细胞核染色时，推荐的Hoechst 33258工作浓度为0.5-10μg/ml。
配制母液
Hoechst 33258溶于水，溶解度可达10mg/ml。因此可配制成10mg/ml的母液于-20℃避光保存
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。
使用方法
1. 用PBS或合适的缓冲液制备10～50µM Hoechst 33258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Hoechst33258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。
a) 对于贴壁细胞或组织切片：加入适量Hoechst 33258染色液，覆盖住样品即可。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。
b) 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。
c) 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350nm，发射波长460nm。
3. 活细胞或组织染色：
a) 细胞培养物中加入适量Hoechst 33258染色液，约1/10细胞培养基体积，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100μl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350nm，发射波长460nm。
备注：产品信息可能会有优化升级。请以实际标签信息为准。
Remark：These protocols are for reference only. Solarbio does not independently validate these methods.
Note：
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2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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