Hoechst 33258 trihydrochloride
(Synonyms: 赫斯特荧光染料33258,bisBenzimide H 33258 trihydrochloride) 目录号 : GC12187
The nucleic acid stain Hoechst 33258 trihydrochloride (Ex/Em: 352/461 nm) is frequently utilized as a cell-permeable nuclear counterstain that emits a blue fluorescence upon binding to dsDNA.
Cas No.:23491-45-4
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
1. Preparing Hoechst 33258 trihydrochloride dye stock solution
1.1 Prepare the Hoechst 33258 trihydrochloride dye stock solution by dissolving the contents of one vial (100 mg) in 10 mL of ddH2O to create a 10 mg/mL (18.7 mM) solution.
Note: The 10 mg/mL Hoechst stock solution may be stored at 2–6°C for up to 1-3 months or at ≤ –20°C for longer periods and should be away from light always.
2. Labeling cells
2.1 Culture cells in an appropriate medium and vessel for fluorescence microscopy.
2.2 Prepare the Hoechst staining solution by diluting the Hoechst stock solution 1:1,000 in PBS.
2.3 Remove the medium in the cell plates.
2.4 Add sufficient staining solution to cover the cells.
2.5 Incubate for 5–10 minutes, protected from light.
2.6 Optional: You may image directly in the staining solution, if you wish.
2.7 Remove the staining solution.
2.8 Wash the cells 3 times in PBS, 5 mins each time.
2.9 Image the cells, Excitation/Emission (nm): 352/461.
This protocol only provides a guideline, and should be modified according to your specific needs.
The nucleic acid stain Hoechst 33258 trihydrochloride (Ex/Em: 352/461 nm) is frequently utilized as a cell-permeable nuclear counterstain that emits a blue fluorescence upon binding to dsDNA. Hoechst 33258 trihydrochloride is commonly employed in various studies related to cell counting, cell cycle analysis, and cell replication. It is particularly useful in identifying condensed nuclei in apoptotic cells, as well as in combination with BrdU staining for cell-cycle studies. Hoechst 33258 works similarly to Hoechst 33342 but is less cell permeable.
Hoechst dyes are also useful for monitoring cell viability by tracking changes in their emission spectra. As minor groove-binding DNA stains with AT selectivity, the Hoechst dyes are able to bind to all nucleic acids, but they show a greater fluorescence enhancement for AT-rich double-stranded DNA strands compared to GC-rich strands [1]. This property has been exploited to identify Q-bands in chromosomes, which are regions rich in AT base pairs that fluoresce brightly when stained with the quinacrine dye [2].
Fig. Fluorescence excitation and emission spectra of Hoechst 33258 bound to DNA
核酸染料Hoechst 33258 trihydrochloride(Ex/Em:352/461 nm)通常作为可穿透细胞膜的核染色剂,在结合dsDNA后发出蓝色荧光,被广泛用于与细胞计数、细胞周期分析和细胞复制相关的各种研究中。它特别适用于鉴定凋亡细胞中的浓缩核,以及与BrdU染色结合用于细胞周期研究。Hoechst 33258的作用类似于Hoechst 33342,但其细胞穿透性较低。
Hoechst染料还可通过跟踪其发射光谱变化来监测细胞存活率。作为具有AT选择性的小沟结合DNA染料,Hoechst染料能够结合所有核酸,但相对于GC丰富的链,它们对富含AT的双链DNA链显示出更大的荧光增强作用[1]。这一特性已被用于鉴定染色体中富含AT碱基对的Q带区域,当用quinacrine染料染色时,这些区域会发出明亮的荧光[2]。
References:
[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 (BBA)-Gene Structure and Expression. 1988 Feb 28;949(2):158-68.
[2]. Weisblum B, Haenssler E. Fluorometric properties of the bibenzimidazole derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma. 1974 Sep;46(3):255-60.
| Cas No. | 23491-45-4 | SDF | |
| 别名 | 赫斯特荧光染料33258,bisBenzimide H 33258 trihydrochloride | ||
| 化学名 | 4-(6-(4-methylpiperazin-1-yl)-1H,1'H-[2,5'-bibenzo[d]imidazol]-2'(3'H)-ylidene)cyclohexa-2,5-dienone trihydrochloride | ||
| Canonical SMILES | CN1CCN(CC1)C2=CC3=C(C=C2)N=C(N3)C4=CC5=C(C=C4)N/C(N5)=C6C=CC(C=C/6)=O.Cl.Cl.Cl | ||
| 分子式 | C25H27Cl3N6O | 分子量 | 533.88 |
| 溶解度 | Water : ≥ 72.85 mg/mL (136.45 mM), DMSO : ≥100 mg/mL | 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.8731 mL | 9.3654 mL | 18.7308 mL |
| 5 mM | 0.3746 mL | 1.8731 mL | 3.7462 mL |
| 10 mM | 0.1873 mL | 0.9365 mL | 1.8731 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。