Hoechst 33258 UltraPure 超纯级

产品介绍

1. 外观：黄色粉末

2. 纯度：≥99% (HPLC) 超纯级

3. 产品参数：Ex(nm) 352   Em(nm) 461   Solvent: Water

产品描述

Hoechst 33258是可透过细胞膜并对DNA染色的细胞核染色试剂，在嵌入双链DNA后释放强烈的蓝色荧光。Hoechst 33258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-DNA的激发和发射波长分别为352 nm和461 nm。 

使用方法

1、对于固定的细胞或组织：

1) 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。

2) 对于贴壁细胞或组织切片，加入少量Hoechst 33258染色液，覆盖住样品即可。对于悬浮细胞，至少加入待染色样品体积3倍的染色液，并混匀。室温放置3-5分钟。

3) 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。

4) 直接在荧光显微镜下或封片后荧光显微镜下观察。观察细胞凋亡时，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：

5) 加入适当量Hoechst 33258染色液，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染色液，对于96 孔板一个孔需加入100微升染色液。

6) 在适宜于细胞培养的温度培养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。 

储存条件：-20℃避光保存，有效期一年。

注意事项

1)Hoechst 33258对人体有一定刺激性，请注意适当防护。

2)荧光染料都存在淬灭的问题，建议染色后尽量当天完成检测。

3)为减缓荧光淬灭可以使用抗荧光淬灭封片液。

4)为了您的安全和健康，请穿实验服并戴一次性手套操作。

Product introduction

1. Appearance: yellow powder

2. Purity: ≥99% (HPLC) ultra-pure grade

3. Product parameters: Ex(nm) 352  Em(nm) 461  Solvent: Water

Product description

Hoechst 33258 is a nuclear staining reagent that can penetrate cell membranes and stain DNA. It releases strong blue fluorescence after intercalating double-stranded DNA. Hoechst 33258 is often used for cell apoptosis detection. After staining, observe with a fluorescence microscope or detect with a flow cytometer. The excitation and emission wavelengths of Hoechst 33258-DNA are 352 nm and 461 nm, respectively.

Instructions

1. For fixed cells or tissues

1) For cell or tissue samples, after fixation, wash them appropriately to remove fixative. Then, if immunofluorescence staining is required, perform immunofluorescence staining first, and then follow the subsequent steps for Hoechst 33258 staining after the staining is completed. If no other staining is required, the subsequent Hoechst 33258 staining is directly performed.

2) For adherent cells or tissue sections, add a small amount of Hoechst 33258 staining solution to cover the sample. For suspended cells, add at least 3 times the volume of the sample to be stained with staining solution and mix well. Leave at room temperature for 3-5 minutes.

3) Aspirate the Hoechst 33258 staining solution, wash 2-3 times with TBST, PBS or saline, 3-5 minutes each time.

4) Observe directly under a fluorescence microscope or after mounting the slide. When observing cell apoptosis, the nucleus of apoptotic cells will be densely stained, or fragmented and densely stained.

2. For living cells or tissues

5) Add an appropriate amount of Hoechst 33258 staining solution, which must fully cover the sample to be stained. Usually, 1ml staining solution is needed for one well of a six-well plate, and 100 microliters of staining solution is required for one well of a 96-well plate.

6) Incubate for 20-30 minutes at a temperature suitable for cell culture. Discard the staining solution, wash 2-3 times with PBS or culture solution to perform fluorescence detection.

Storage conditions: Store at -20°C and protected from light, valid for one year.

Precautions

1) Hoechst 33258 is irritating to the human body, please pay attention to proper protection.

2) Fluorescent dyes have the problem of quenching. It is recommended to complete the test on the same day after dyeing.

3) To slow down fluorescence quenching, anti-fluorescence quenching mounting solution can be used.

4) For your safety and health, please wear lab coats and disposable gloves for operation.

Hoechst 33342 UltraPure 超纯级

产品介绍

1. 外观：黄绿色粉末

2. 纯度：≥99% (HPLC) 超纯级

3. 产品参数：Ex(nm) 350    Em(nm) 461    Solvent: Water

产品描述

Hoechst染料是一类在显微观察中标记DNA的荧光染料。因为这类荧光染料能标记DNA，所以它们也经常用于细胞核和线粒体的显像观察。这类染料中两个相关的染料Hoechst 33258和Hoechst 33342经常使用。这两种染料都在紫外光下350nm处被激发，都在461nm处最大发射光附近发射蓝/青色荧光。Hoechst染料可以用于活细胞或者固定化细胞，并且经常用来代替其它核酸染料如DAPI。这两种染料关键的不同点在于，Hoechst 33342加有乙基，这使它具有更强的亲脂性，因此能更好的透过完整的细胞膜。在一些实验中，Hoechst 33258的渗透性明显比Hoechst 33342要弱些。这些染料也可以用来检测样品中的DNA含量，通过绘制发射光强度与DNA含量的标准曲线。

Hoechst 33342是一种可透过细胞膜并对DNA染色的细胞核染色试剂，它在嵌入双链DNA后释放强烈的蓝色荧光。Hoechst 33342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 33342-DNA的激发和发射波长分别为350 nm和460 nm。Hoechst 33342溶于水，溶解度可达20mg/mL。

染色程序

（1） 用PBS或合适的缓冲液制备10～50μM Hoechst33342染料。

（2） 将1/10细胞培养基体积的Hoechst染料溶液加入细胞培养物中（可以用1/10浓度的Hoechst染料缓冲液代替培养基）。

（3）在37℃培养细胞10～20分钟。

（4）用PBS或合适的缓冲液洗细胞两次。

（5）用带有350nm激发波长，460nm发射波长的滤光片的荧光显微镜观察细胞。

储存条件：-20℃干燥避光保存，有效期一年。

注意事项

1.Hoechst 33342对人体有一定刺激性，请注意适当防护。

2.荧光染料都存在淬灭的问题，建议染色后尽量当天完成检测。

3.为减缓荧光淬灭可以使用抗荧光淬灭封片液。

4.为了您的安全和健康，请穿实验服并戴一次性手套操作。

Product introduction

1. Appearance: yellow-green powder

2. Purity: ≥99% (HPLC) ultra-pure grade

3. Product parameters: Ex(nm) 350 Em(nm) 461 Solvent: Water

Product description

Hoechst dyes are a class of fluorescent dyes that mark DNA in microscopic observation. Because such fluorescent dyes can label DNA, they are also often used for imaging and observation of cell nuclei and mitochondria. Two related dyes of this type, Hoechst 33258 and Hoechst 33342, are frequently used. Both dyes are excited at 350nm under ultraviolet light, and both emit blue/cyan fluorescence near the maximum emission at 461nm. Hoechst dyes can be used in living or immobilized cells, and are often used to replace other nucleic acid dyes such as DAPI. The key difference between the two dyes is that Hoechst 33342 is added with ethyl groups, which makes it more lipophilic and therefore can better penetrate the complete cell membrane. In some experiments, the permeability of Hoechst 33258 is obviously weaker than that of Hoechst 33342. These dyes can also be used to detect the DNA content in the sample by drawing a standard curve between the emitted light intensity and the DNA content.

Hoechst 33342 is a nuclear staining reagent that can penetrate cell membranes and stain DNA. It releases strong blue fluorescence after intercalating double-stranded DNA. Hoechst 33342 is often used for apoptosis detection. After staining, it is observed with a fluorescence microscope or detected by flow cytometry. The excitation and emission wavelengths of Hoechst 33342-DNA are 350 nm and 460 nm, respectively. Hoechst 33342 is soluble in water with a solubility of up to 20mg/mL.

Dyeing procedure

(1) Prepare 10-50μM Hoechst33342 dye with PBS or a suitable buffer.

(2) Add 1/10 of the cell culture medium volume of Hoechst dye solution to the cell culture (1/10 concentration of Hoechst dye buffer can be used to replace the medium).

(3) Incubate the cells at 37°C for 10-20 minutes.

(4) Wash the cells twice with PBS or a suitable buffer.

(5) Observe the cells with a fluorescence microscope with a 350nm excitation wavelength and a 460nm emission wavelength filter.

Storage conditions: Store at -20°C dry and dark, valid for one year.

Precautions

1. Hoechst 33342 is irritating to the human body, please pay attention to proper protection.

2. Fluorescent dyes have the problem of quenching. It is recommended to complete the test on the same day after dyeing.

3. To slow down fluorescence quenching, anti-fluorescence quenching mounting solution can be used.

4. For your safety and health, please wear lab coats and disposable gloves for operation.