Hoechst 33258 (Synonyms: bisBenzimide H 33258; H 33258)

目录号: HY-15558 纯度: 99.11%: Data Sheet SDS COA 产品使用指南
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Hoechst 33258 是一种蓝色至蓝绿色荧光的活细胞染料，可标记 DNA。Hoechst 33258 可特异性结合 DNA 小沟 (同时倾向于结合富含 A/T 的 DNA)，使荧光强度显著增强。Hoechst 33258 可穿过细胞膜，导致 DNA 结构变化，如 G2/M 期阻滞。Hoechst 33258 可结合活细胞或固定细胞，荧光强度随着溶液 pH 升高而增强。Hoechst 33258 作为 DNA 特异性探针，可用于检测 DNA 含量、分析细胞周期等。Hoechst 33258 的激发波长为 350-365 nm，发射波长为 460-490 nm。

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Hoechst 33258 Chemical Structure

CAS No. : 23491-44-3

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Other Forms of Hoechst 33258:

MCE 顾客使用本产品发表的 17 篇科研文献

: Hoechst 33258 purchased from MCE. Usage Cited in: Virulence. 2022 Dec;13(1):444-457. [Abstract]; The apoptotic rate was detected by Hoechst staining (400×), HeLa cells were treated with siRNA for 24 h and followed by persistent infection.

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生物活性
实验参考方法
纯度 & 产品资料
参考文献

生物活性

Hoechst 33258 is a blue to blue-green fluorescent live cell dye that can label DNA. Hoechst 33258 can specifically bind to the minor groove of DNA (and tends to bind to A/T-rich DNA), resulting in a significant increase in fluorescence intensity. Hoechst 33258 can cross the cell membrane and cause changes in DNA structure, such as G2/M phase arrest. Hoechst 33258 can bind to live or fixed cells, and the fluorescence intensity increases with increasing solution pH. As a DNA-specific probe, Hoechst 33258 can be used to detect DNA content, analyze cell cycle, etc. The excitation wavelength of Hoechst 33258 is 350-365 nm, and the emission wavelength is 460-490 nm^[1]^[2]^[3].

细胞效力
(Cellular Effect)

Cell Line	Type	Value	Description	References
A2780	IC₅₀	12 μM Compound: HOE-33258	Antitumor activity of compound for 96-h exposure in A2780 human ovarian cell line Antitumor activity of compound for 96-h exposure in A2780 human ovarian cell line	[PMID: 11170623]
A2780cisR	IC₅₀	9.5 μM Compound: HOE-33258	Antitumor activity of compound for 96-h exposure in A2780cisR (acquired resistance to cisplatin) human ovarian cell line Antitumor activity of compound for 96-h exposure in A2780cisR (acquired resistance to cisplatin) human ovarian cell line	[PMID: 11170623]
A549	IC₅₀	≥ 31.2 μg/mL Compound: 2	Cytotoxicity against human A549 cells assessed as reduction in cell survival after 24 hrs by resazurin assay Cytotoxicity against human A549 cells assessed as reduction in cell survival after 24 hrs by resazurin assay	[PMID: 27301676]
AGS	GI₅₀	112 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human AGS cells after 72 hrs by MTT assay Cytotoxicity against human AGS cells after 72 hrs by MTT assay	[PMID: 21227551]
BEAS-2B	IC₅₀	≥ 31.2 μg/mL Compound: 2	Cytotoxicity against human BEAS2B cells assessed as reduction in cell survival after 24 hrs by resazurin assay Cytotoxicity against human BEAS2B cells assessed as reduction in cell survival after 24 hrs by resazurin assay	[PMID: 27301676]
Caco-2	GI₅₀	84 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human Caco2 cells after 72 hrs by MTT assay Cytotoxicity against human Caco2 cells after 72 hrs by MTT assay	[PMID: 21227551]
CH1	IC₅₀	26.5 μM Compound: HOE-33258	Antitumor activity of compound for 96-h exposure in CH1 human ovarian cell line Antitumor activity of compound for 96-h exposure in CH1 human ovarian cell line	[PMID: 11170623]
HeLa	GI₅₀	115.7 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human HeLa cells after 72 hrs by MTT assay Cytotoxicity against human HeLa cells after 72 hrs by MTT assay	[PMID: 21227551]
HeLa	IC₅₀	51.31 μM Compound: H. 33258, Hoechst 33258	Cytotoxicity against human HeLa cells after 72 hrs by MTT assay Cytotoxicity against human HeLa cells after 72 hrs by MTT assay	[PMID: 22959518]
HEp-2	GI₅₀	132.5 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human Hep2 cells after 72 hrs by MTT assay Cytotoxicity against human Hep2 cells after 72 hrs by MTT assay	[PMID: 21227551]
HL-60	IC₅₀	32.43 μM Compound: H. 33258, Hoechst 33258	Cytotoxicity against human HL60 cells after 72 hrs by MTT assay Cytotoxicity against human HL60 cells after 72 hrs by MTT assay	[PMID: 22959518]
MCF7	GI₅₀	5.7 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay Cytotoxicity against human MCF7 cells after 72 hrs by MTT assay	[PMID: 21227551]
MDCK	GI₅₀	157.5 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against MDCK cells after 72 hrs by MTT assay Cytotoxicity against MDCK cells after 72 hrs by MTT assay	[PMID: 21227551]
MIA PaCa-2	GI₅₀	191.5 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human MIAPaCa2 cells after 72 hrs by MTT assay Cytotoxicity against human MIAPaCa2 cells after 72 hrs by MTT assay	[PMID: 21227551]
NCI-H358	GI₅₀	91.7 x 10^-6 mol/dm³ Compound: Hoechst 33258	Cytotoxicity against human NCI-H358 cells after 72 hrs by MTT assay Cytotoxicity against human NCI-H358 cells after 72 hrs by MTT assay	[PMID: 21227551]
SK-OV-3	IC₅₀	> 100 μM Compound: HOE-33258	Antitumor activity of compound for 96-h exposure in SKOV-3 human ovarian cell line Antitumor activity of compound for 96-h exposure in SKOV-3 human ovarian cell line	[PMID: 11170623]
U-937	IC₅₀	15.42 μM Compound: H. 33258, Hoechst 33258	Cytotoxicity against human U937 cells after 72 hrs by MTT assay Cytotoxicity against human U937 cells after 72 hrs by MTT assay	[PMID: 22959518]

体外研究
(In Vitro)

Hoechst 33258 有两种结合模式：Type 1 和 Type 2，Type 1 在低浓度下结合，荧光量子产率高，发射峰在 460 nm；Type 2 在较高浓度下结合，导致荧光猝灭，发射峰在 490 nm 左右^[1]。
Hoechst 33258 对 HeLa、HL60、U937 细胞的 IC₅₀ 分别为 51.31 μM、32.43 μM、15.42 μM^[2]。

Hoechst 33258 的使用方法参考^[1]
1 Hoechst 工作液的配制：
1.1 储备液配制将 Hoechst 33258 粉末配制成 1-10 mM 储备液，避光搅拌至完全溶解。
Note：染料对光敏感，配制过程需避光。
1.2 工作液稀释
使用前用 PBS 或细胞培养基将储备液稀释至工作浓度，通常为 5-10 μg/mL (细胞染色) 或 1-50 μM (DNA 结合研究) 。
Note：现用现配以避免荧光淬灭。
2 贴壁细胞染色步骤：
将细胞接种于培养板/皿中，培养至对数生长期。如需固定细胞，用 4% 多聚甲醛室温固定 10-15 min，PBS 洗涤 2 次，每次 5 分钟。
2.1 加入 Hoechst 33258 工作液，37°C 避光孵育 10-30 min (根据细胞类型调整时间，避免过度染色) 。
2.2 对于活细胞染色，直接加入培养基中孵育，无需固定；对于死细胞，可先固定再染色。
2.3 用 PBS或无血清培养基洗涤细胞 2-3 次，去除未结合的染料，减少背景荧光。
3 悬浮细胞染色步骤：
3.1 收集细胞，1000 rpm 离心 5 min，弃上清。
3.2 用 PBS 重悬细胞，加入 Hoechst 33258 工作液，室温避光孵育 15-20 min。
3.3 离心洗涤后重悬，用于流式细胞术或荧光显微镜检测。
4 检测通道与工具：
4.1 激发波长 (Ex) ：350-365 nm (紫外光或近紫外光) 。
4.2 发射波长 (Em) ：460-490 nm (蓝色至蓝绿色荧光) 。
4.3 检测工具：
荧光显微镜：配备紫外激发滤光片 (如 DAPI 通道) ，用于观察细胞内 DNA 分布。
流式细胞仪：使用紫外激光器 (355 nm) 和 450/50 nm 带通滤光片，检测细胞周期或 DNA 含量。
酶标仪：多孔板检测时，选用紫外激发和蓝色发射通道，用于高通量 DNA 定量。
5 注意事项：
5.1 需根据细胞类型和实验目的调整浓度，高浓度 (>50 μM) 可能导致 DNA 沉淀 (如，D/P>0.4 时沉淀) 。
5.2 短期孵育 (<30 min) 对细胞毒性较低，长期孵育需验证细胞活力。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

分子量

424.50

Formula

C₂₅H₂₄N₆O

CAS 号

23491-44-3

性状

固体

颜色

Light yellow to yellow

Emission (Em)

451

Excitation (Ex)

356

运输条件

Room temperature in continental US; may vary elsewhere.

储存方式

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

溶解性数据

细胞实验：

DMSO 中的溶解度 : 50 mg/mL (117.79 mM; 超声助溶; 吸湿的 DMSO 对产品的溶解度有显著影响，请使用新开封的 DMSO)

配制储备液

浓度溶剂体积质量	1 mg	5 mg	10 mg

1 mM	2.3557 mL	11.7786 mL	23.5571 mL
5 mM	0.4711 mL	2.3557 mL	4.7114 mL
10 mM	0.2356 mL	1.1779 mL	2.3557 mL

查看完整储备液配制表

* 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, 6 months; -20°C, 1 month (protect from light)。-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。

摩尔计算器
稀释计算器

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

浓度 _(start)

体积 _(start)

浓度 _(final)

体积 _(final)

动物实验：

请根据您的实验动物和给药方式选择适当的溶解方案。

以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：
——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用；
以下溶剂前显示的百分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

方案一

请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.08 mg/mL (4.90 mM); 澄清溶液

此方案可获得 ≥ 2.08 mg/mL（饱和度未知）的澄清溶液。
以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；再向上述体系中加入 50 μL Tween-80，混合均匀；然后再继续加入 450 μL 生理盐水定容至 1 mL。
生理盐水的配制：将 0.9 g 氯化钠，溶解于 ddH₂O 并定容至 100 mL，可以得到澄清透明的生理盐水溶液。
方案二

请依序添加每种溶剂： 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.08 mg/mL (4.90 mM); 澄清溶液

此方案可获得 ≥ 2.08 mg/mL（饱和度未知）的澄清溶液。
以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。
2 g SBE-β-CD（磺丁基醚 β-环糊精）粉末定容于 10 mL 的生理盐水中，完全溶解至澄清透明。

动物溶解方案计算器

请输入动物实验的基本信息：

给药剂量

mg/kg

动物的平均体重

每只动物的给药体积

μL

动物数量

只

由于实验过程有损耗，建议您多配一只动物的量

请输入您的动物体内配方组成：

DMSO +

Tween-80 +

Saline

如果您的动物是免疫缺陷鼠或者体弱鼠，建议 DMSO 中的在最后工作液体系中的占比尽量不超过 2%。

方案所需 助溶剂 包括：DMSO，，均可在 MCE 网站选购。，Tween 80，均可在 MCE 网站选购。

计算结果

工作液所需浓度 : mg/mL

储备液配制方法 : mg 药物溶于 μL DMSO（母液浓度为 mg/mL）。

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

您所需的储备液浓度超过该产品的实测溶解度，以下方案仅供参考，如有需要，请与 MCE 中国技术支持联系。

动物实验体内工作液的配制方法 : 取 μL DMSO 储备液，加入 μL 。 μL ，混合均匀至澄清，再加 μL Tween 80，混合均匀至澄清，再加 μL 生理盐水。

连续给药周期超过半月以上，请谨慎选择该方案。

请确保第一步储备液溶解至澄清状态，从左到右依次添加助溶剂。您可采用超声加热（超声清洗仪，建议频次 20-40 kHz），涡旋吹打等方式辅助溶解。

纯度 & 产品资料

纯度: 99.93%

选择批次：

Data Sheet (666 KB) SDS (393 KB)

COA (283 KB) LCMS (134 KB)

产品使用指南 (1538 KB)

染色示例

参考文献

[1]. Stokke T, et al. Multiple binding modes for Hoechst 33258 to DNA. J Histochem Cytochem. 1985 Apr;33(4):333-8. [Content Brief]

[2]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through induction of apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [Content Brief]

[3]. Pjura PE, et al. Binding of Hoechst 33258 to the minor groove of B-DNA. J Mol Biol. 1987 Sep 20;197(2):257-71. [Content Brief]

Cell Assay ^[2]	Hoechst 33258 is prepared as stock solutions in highly pure water. Working solutions in a concentration range of 10^-3-10^-6 mol/dm³ are prepared prior to testing. Cytotoxic effects of Hoechst 33258 on tested cell lines are determined by the MTT assay. Cells are seeded in 96 micro well flat bottom plates at a concentration of 2×10⁴ cells/mL and left overnight in the CO₂ incubator allowing them to attach to the plate surface. Growing medium is replaced with compound supplemented or control medium and incubated for 72 h. Fresh medium with 5 mg/mL of MTT is added onto cells and incubated for 4 h at 37°C. Upon media removal, water insoluble MTT-formazan crystals formed inside the living cells are dissolved in DMSO and the absorbance at 570 nm proportional to the number of living cells is measured on an Elisa Microplate Reader. All experiments are performed at least three times in triplicates.The GI₅₀ value, defined as the compound concentration (μM) leading to cellular growth inhibition by 50%, is calculated and used as a parameter to compare cytotoxicity among the compounds^[2]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献	[1]. Stokke T, et al. Multiple binding modes for Hoechst 33258 to DNA. J Histochem Cytochem. 1985 Apr;33(4):333-8. [Content Brief] [2]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through induction of apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [Content Brief] [3]. Pjura PE, et al. Binding of Hoechst 33258 to the minor groove of B-DNA. J Mol Biol. 1987 Sep 20;197(2):257-71. [Content Brief]

Cell Assay
^[2]

Hoechst 33258 is prepared as stock solutions in highly pure water. Working solutions in a concentration range of 10^-3-10^-6 mol/dm³ are prepared prior to testing. Cytotoxic effects of Hoechst 33258 on tested cell lines are determined by the MTT assay. Cells are seeded in 96 micro well flat bottom plates at a concentration of 2×10⁴ cells/mL and left overnight in the CO₂ incubator allowing them to attach to the plate surface. Growing medium is replaced with compound supplemented or control medium and incubated for 72 h. Fresh medium with 5 mg/mL of MTT is added onto cells and incubated for 4 h at 37°C. Upon media removal, water insoluble MTT-formazan crystals formed inside the living cells are dissolved in DMSO and the absorbance at 570 nm proportional to the number of living cells is measured on an Elisa Microplate Reader. All experiments are performed at least three times in triplicates.The GI₅₀ value, defined as the compound concentration (μM) leading to cellular growth inhibition by 50%, is calculated and used as a parameter to compare cytotoxicity among the compounds^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

参考文献

[1]. Stokke T, et al. Multiple binding modes for Hoechst 33258 to DNA. J Histochem Cytochem. 1985 Apr;33(4):333-8. [Content Brief]

[2]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through induction of apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [Content Brief]

[3]. Pjura PE, et al. Binding of Hoechst 33258 to the minor groove of B-DNA. J Mol Biol. 1987 Sep 20;197(2):257-71. [Content Brief]

Hoechst 33258 相关分类

Others

完整储备液配制表

可选溶剂	浓度溶剂体积质量	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	2.3557 mL	11.7786 mL	23.5571 mL	58.8928 mL
	5 mM	0.4711 mL	2.3557 mL	4.7114 mL	11.7786 mL
	10 mM	0.2356 mL	1.1779 mL	2.3557 mL	5.8893 mL
	15 mM	0.1570 mL	0.7852 mL	1.5705 mL	3.9262 mL
	20 mM	0.1178 mL	0.5889 mL	1.1779 mL	2.9446 mL
	25 mM	0.0942 mL	0.4711 mL	0.9423 mL	2.3557 mL
	30 mM	0.0785 mL	0.3926 mL	0.7852 mL	1.9631 mL
	40 mM	0.0589 mL	0.2945 mL	0.5889 mL	1.4723 mL
	50 mM	0.0471 mL	0.2356 mL	0.4711 mL	1.1779 mL
	60 mM	0.0393 mL	0.1963 mL	0.3926 mL	0.9815 mL
	80 mM	0.0294 mL	0.1472 mL	0.2945 mL	0.7362 mL
	100 mM	0.0236 mL	0.1178 mL	0.2356 mL	0.5889 mL

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Hoechst 33258 (Synonyms: bisBenzimide H 33258; H 33258)

Customer Review

Other Forms of Hoechst 33258:

MCE 顾客使用本产品发表的 17 篇科研文献

Hoechst 33258 相关产品

•同靶点产品:

•同靶点蛋白产品:

生物活性

实验参考方法

纯度 & 产品资料

参考文献

摩尔计算器

稀释计算器

Hoechst 33258 相关分类

完整储备液配制表

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Hoechst 33258 (Synonyms: bisBenzimide H 33258; H 33258)

Customer Review

Other Forms of Hoechst 33258:

Hoechst 33258 相关抗体

MCE 顾客使用本产品发表的 17 篇科研文献

Hoechst 33258 相关产品

•同靶点产品:

•同靶点蛋白产品:

生物活性

实验参考方法

纯度 & 产品资料

参考文献

Hoechst 33258 相关抗体:

摩尔计算器

稀释计算器

Hoechst 33258 相关分类

完整储备液配制表

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Request for HNMR Report

Request for HNMR Report

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