Hoechst 33258 Stain solution(1mg/ml)
Cat.No:C0021 Solarbio
Storage:Store at -20℃,avoid light,1 year
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Name | Hoechst 33258 Stain solution(1mg/ml) |
Storage | Store at -20℃,avoid light,1 year |
Unit | Kit |
Specification | 1ml 10*1ml |
Hoechst 33258,也称bisBenzimide H 33258或HOE 33258,是一种可以穿透细胞膜的蓝色荧光染料,对细胞的毒性较低。Hoechst 33258染色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也常用于普通的细胞核染色,或常规的DNA染色。Hoechst 33258的最大激发波长为346nm,最大发射波长为460nm;Hoechst 33258和双链DNA结合后,最大激发波长为352nm,最大发射波长为461nm。本Hoechst 33258染色液可直接用于固定细胞或组织的细胞核染色,也可直接用于活细胞或组织的细胞核染色。
使用说明:
使用前用生理盐水或PBS将Hoechst 33258染色液稀释100倍,即为工作液。
1. 对于固定的细胞或组织:
a. 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。
b. 对于贴壁细胞或组织切片,加入少量Hoechst 33258工作液,覆盖住样品即可;对于悬浮细胞,至少加入待染色样品3倍体积的工作液,混匀。室温放置3-5分钟。
c. 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。
d. 直接在荧光显微镜下观察或封片后荧光显微镜下观察。细胞发生凋亡时,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:
a. 加入适当量Hoechst 33258工作液,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工作液,对于96孔板一个孔需加入100μL工作液。
b. 在适宜于细胞培养的温度下培养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。
注意事项:
荧光染料都存在淬灭的问题,为减缓荧光淬灭可以使用抗荧光衰减封片剂。建议染色后尽量当天完成检测,活细胞或组织染色后应立即观察。
为了您的安全和健康,请穿实验服并戴一次性手套操作。
Remark:These protocols are for reference only. Solarbio does not independently validate these methods.
Note:
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
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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