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细胞相关> 细胞死亡与自噬> 细胞凋亡
细胞凋亡-Hoechst染色试剂盒
产品编号: C0003
产品包装:100次
选择包装
价格:¥388.00元
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
C0003 细胞凋亡-Hoechst染色试剂盒 100次 388.00元

碧云天生产的细胞凋亡-Hoechst染色试剂盒(Hoechst Staining Kit)为您提供了一种经典而又快速简便的细胞凋亡检测方法。细胞发生凋亡时,染色质会固缩。所以Hoechst 33258染色后,在荧光显微镜下观察,正常细胞的细胞核呈正常的蓝色,而凋亡细胞的细胞核会呈致密浓染,或呈碎块状致密浓染,颜色有些发白。典型的Hoechst 33258染色后的细胞图片参考下图。


正常细胞核 刺激后有致密浓染的凋亡细胞

只需25分钟即可完成细胞凋亡检测的整个过程。
本试剂盒提供了固定液,染色液,及抗荧光淬灭封片液。
本试剂盒对贴壁细胞,悬浮细胞和组织切片均适用。
足够检测100个样品。
包装清单:

产品编号 产品名称 包装
C0003-1 固定液 50ml
C0003-2 Hoechst 33258染色液 50ml
C0003-3 抗荧光淬灭封片液 5ml
说明书 1份

保存条件:
4℃保存,Hoechst 33258染色液需4℃避光保存,半年有效。
注意事项:
需可以观察蓝色荧光的荧光显微镜或激光共聚焦显微镜。
需PBS或0.9%NaCl溶液。
需自备盖玻片与载玻片。盖玻片载玻片可以向碧云天订购。
荧光物质均易发生淬灭,染色后的样品宜避光保存。可以向碧云天选购各种型号的载玻片存储盒
在使用抗淬灭封片液的情况下可以减缓淬灭,但仍宜尽量避光。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
1.贴壁细胞
a.取洁净盖玻片在70%乙醇中浸泡5分钟或更长时间,无菌超净台内吹干或用无菌的PBS或0.9%NaCl等溶液洗涤三遍,再用细胞培养液洗涤一遍。将盖玻片置于六孔板内,种入细胞培养过夜,使约为50%-80%满。
b.刺激细胞发生凋亡后,吸尽培养液,加入0.5ml 固定液,固定10分钟或更长时间(可4℃过夜)。
c.去固定液,用PBS或0.9%NaCl 洗两遍,每次3分钟,吸尽液体。洗涤时宜用摇床,或手动晃动。
d.加入0.5ml Hoechst 33258染色液,染色5分钟。也宜用摇床,或手动晃动数次。
e.去染色液,用PBS或0.9%NaCl 洗两遍,每次3分钟,吸尽液体。洗涤时宜用摇床,或手动晃动。
f.滴一滴抗荧光淬灭封片液于载玻片上,盖上贴有细胞的盖玻片,让细胞接触封片液,尽量避免气泡。
g.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右,发射波长460nm左右,图谱请参考下图。
2.悬浮细胞
a.离心收集细胞样品于1.5ml离心管内,加入0.5ml 固定液,缓缓悬起细胞,固定10分钟或更长时间(可4℃过夜)。
b.离心去固定液,用PBS或0.9%NaCl 洗两遍,每次3分钟。洗涤期间手动晃动数次。
c.离心后吸去大部分液体保留约50μl液体,再缓缓悬起细胞,滴加至载玻片上,尽量使细胞分布均匀。
d.稍晾干,使细胞贴在载玻片上不易随液体流动。
e.均匀滴上0.5ml Hoechst 33258染色液,染色5分钟。用吸水纸从边缘吸去液体,微晾干。
f.去染色液,用PBS或0.9%NaCl 洗两遍,每次3分钟,吸尽液体。洗涤时宜用摇床,或手动晃动。
g.滴一滴抗荧光淬灭封片液于载玻片上,盖上一洁净的盖玻片,尽量避免气泡。
h.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右,发射波长460nm左右,图谱请参考下图。
3.组织切片
a.常规包埋切片后,根据切片的不同类型,处理至可以用于免疫组化染色。
b.PBS或0.9%NaCl 洗两遍,每次3分钟,吸尽液体。洗涤时宜用摇床,或手动晃动。可在六孔板中操作。
c.加入0.5ml Hoechst 33258染色液,染色5分钟。也宜用摇床,或手动晃动数次。
d.去染色液,用PBS或0.9%NaCl 洗两遍,每次3分钟,吸尽液体。洗涤时宜用摇床,或手动晃动。
e.小心将切片置于载玻片上,滴一滴抗淬灭封片液,盖上一洁净的盖玻片,尽量避免气泡。
f.荧光显微镜可检测到呈蓝色的细胞核。激发波长350nm左右,发射波长460nm左右,图谱请参考下图。

                                     Hoechst 33258的吸收光谱和发射光谱,左侧峰为吸收光谱,右侧峰为发射光谱。

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