meta-iodoHoechst 33258
目录号 : GC36586meta-iodoHoechst 33258是核酸染色剂Hoechst 33258的类似物,与dsDNA结合后发出蓝色荧光。
Cas No.:158013-42-4
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导,请根据您的具体需要进行修改。
1、制备Hoechst染色液
(1) 配制Hoechst染料储存液: 使用DMSO溶解固体粉末,配置成10mg/mL的Hoechst染料储存液。
注意: Hoechst储存液建议分装后于-4℃或-20℃避光保存,避免反复冻融。
(2)工作液制备:使用预热的无血清培养基或缓冲液(如HBSS或PBS)稀释储存液,配制浓度为10μg/mL的Hoechst工作液。
注意: 请根据实际情况调整 Hoechst 工作液浓度,现用现配。
2、细胞染色
2.1 悬浮细胞(以6孔板为例)
(1)悬浮细胞经1000g离心3-5min。弃去上清液,使用PBS清洗两次,每次5分钟。
(2)加入1mL的Hoechst染料工作液,室温避光孵育5-10 min分钟。
(3)孵育结束后,经1000g离心5分钟,去除上清液,加入PBS清洗2-3次,每次5分钟。
(4)使用无血清细胞培养基或PBS重悬细胞,通过荧光显微镜或流式细胞技术进行观察。
2.2 贴壁细胞
(1)在无菌盖玻片上培养贴壁细胞。
(2)从培养基中移走盖玻片,吸出过量的培养基,将盖玻片放在潮湿的环境中。
(3)从盖玻片的一角加入100uL的Hoechst染料工作液,轻轻晃动使染料均匀覆盖所有细胞,室温避光孵育5-15min分钟。
(4)吸弃染料工作液,使用培养液洗盖玻片2~3次,通过荧光显微镜进行观察。
注意事项:
①对于固定的细胞或组织样品的染色,固定后需漂洗去除固定剂;
②Hoechst 33258染色通常在其他染色后进行,如果不需要进行其它染色,则直接进行Hoechst 33258染色;
③为减缓荧光淬灭,建议使用抗荧光淬灭封片剂;
④荧光染料均存在淬灭问题,请尽量注意避光;
⑤Hoechst 33258对人体有一定刺激性,为了您的安全和健康,请穿实验服并戴一次性手套操作。
References:
[1]. K D Harshman, P B Dervan. Molecular recognition of B-DNA by Hoechst 33258. 1985 Jul 11;13(13):4825-35. doi: 10.1093/nar/13.13.4825.
Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA.IC50 Value:Target: These Bis-benzimides were originally developed by Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33342nd compound made by the company. There are three related Hoechst stains: Hoechst 33258, Hoechst 33342, and Hoechst 34580. The dyes Hoechst 33258 and Hoechst 33342 are the ones most commonly used and they have similarexcitation/emission spectra. Both dyes are excited by ultraviolet light at around 350 nm, and both emit blue/cyan fluorescent light around anemission maximum at 461 nm. Unbound dye has its maximum fluorescence emission in the 510-540 nm range. Hoechst dyes are soluble in water and in organic solvents such as dimethyl formamide or dimethyl sulfoxide. Concentrations can be achieved of up to 10 mg/mL. Aqueous solutions are stable at 2-6 °C for at least six months when protected from light. For long-term storage the solutions are instead frozen at ≤-20 °C.The dyes bind to the minor groove of double-stranded DNA with a preference for sequences rich in adenine andthymine. Although the dyes can bind to all nucleic acids, AT-rich double-stranded DNA strands enhance fluorescence considerably.Hoechst dyes are cell-permeable and can bind to DNA in live or fixed cells. Therefore, these stains are often called supravital, which means that cells survive a treatment with these compounds. Cells that express specific ATP-binding cassette transporter proteins can also actively transport these stains out of their cytoplasm.in vitro: N/A in vivo: N/AClinical trial: N/A
[1]. Portugal J, Waring MJ. Assignment of DNA binding sites for 4',6-diamidine-2-phenylindole and bisbenzimide (Hoechst 33258). A comparative footprinting study. Biochimica et Biophysica Acta 949 (2): 158-68. [2]. Latt SA, Stetten G, Juergens LA, Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 23 (7): 493-505. [3]. a b c "Hoechst Stains". Invitrogren (Molecular Probes).
Cas No. | 158013-42-4 | SDF | |
Canonical SMILES | CN1CCN(C2=CC=C3N=C(C4=CC=C5N=C(C6=CC=CC(I)=C6)NC5=C4)NC3=C2)CC1 | ||
分子式 | C25H23IN6 | 分子量 | 534.39 |
溶解度 | Soluble in DMSO | 储存条件 | 4°C, protect from light |
General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液;一旦配成溶液,请分装保存,避免反复冻融造成的产品失效。 储备液的保存方式和期限:-80°C 储存时,请在 6 个月内使用,-20°C 储存时,请在 1 个月内使用。 为了提高溶解度,请将管子加热至37℃,然后在超声波浴中震荡一段时间。 |
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Shipping Condition | 评估样品解决方案:配备蓝冰进行发货。所有其他可用尺寸:配备RT,或根据请求配备蓝冰。 |
制备储备液 | |||
1 mg | 5 mg | 10 mg | |
1 mM | 1.8713 mL | 9.3565 mL | 18.7129 mL |
5 mM | 0.3743 mL | 1.8713 mL | 3.7426 mL |
10 mM | 0.1871 mL | 0.9356 mL | 1.8713 mL |
第一步:请输入基本实验信息(考虑到实验过程中的损耗,建议多配一只动物的药量) | ||||||||||
给药剂量 | mg/kg | 动物平均体重 | g | 每只动物给药体积 | ul | 动物数量 | 只 | |||
第二步:请输入动物体内配方组成(配方适用于不溶于水的药物;不同批次药物配方比例不同,请联系GLPBIO为您提供正确的澄清溶液配方) | ||||||||||
% DMSO % % Tween 80 % saline | ||||||||||
计算重置 |
计算结果:
工作液浓度: mg/ml;
DMSO母液配制方法: mg 药物溶于 μL DMSO溶液(母液浓度 mg/mL,
体内配方配制方法:取 μL DMSO母液,加入 μL PEG300,混匀澄清后加入μL Tween 80,混匀澄清后加入 μL saline,混匀澄清。
1. 首先保证母液是澄清的;
2.
一定要按照顺序依次将溶剂加入,进行下一步操作之前必须保证上一步操作得到的是澄清的溶液,可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。