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基本描述
| 英文名称 | DRAQ5 Fluorescent Probe | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| 储存温度 | 2-8°C储存,避光 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 运输条件 | 冰袋运输 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 产品介绍 |
产品介绍 1.?规格：5mM?50?μL（适用4 个?96 孔板的实验）,?200?μL（适用16个?96 孔板的实验） 2.?参数：MW=?412.49；?Wavelength: Ex/Em=?646/697nm；Solvent: Water 3.?储存：4 ℃避光保存2年 产品描述 DRAQ5是一种对双链DNA具有高亲和力的远红外荧光活细胞DNA染料。它是一种可以透过细胞膜的染料，可标记活细胞或固定后/死细胞。在流式细胞术中，这种染料可用于区分有核和无核细胞。由于DRAQ5能够按照化学计量比结合至DNA因此还可用于报告细胞核DNA含量，适用于染色体倍数和细胞周期分析。在荧光显微镜分析中，它可用作细胞核复染剂。DRAQ5有很多应用，高度兼容现有仪器平台广泛使用的程序，主要的应用领域HCS，细胞模型，GFP，流式细胞仪和荧光显微镜。 DRAQ5激发波长范围为488～647nm。对于成像显微术，建议使用633或647nm的光源进行激发。对于流式细胞术，在488nm处激发这种染料时，可使用685LP二向分色镜和710/50通道进行检测；在633nm处激发时， 可以使用660/20通道进行检测。对于细胞周期/DNA?分析应用，建议使用波长较长的滤光片，例如735LP二向分色镜和?780/60通道来优化G1和G2/M峰的CV值。请确保您的仪器能够检测该染料。 由于DRAQ发射和激发波长范围很宽，不建议将?DRAQ5与其他可被488或633nm激发的远红光荧光染料联用。 操作说明 （1）说明 ②.用PBS或培养基重悬细胞，控制细胞密度≤ 4 × 105 cells/mL。对于贴壁细胞和部分组织，大致估计细胞个数。 ③.按下表1加入相应体积的合适浓度的DRAQ5?染色液，?DRAQ5染色液可以直接加到组织或者贴壁细胞的表面，或者直接加入到新鲜培养基中 ④.轻轻混匀，室温下孵育5-30min。37℃ 孵育，时间缩短 为1-3min。对于时间跨度较长的实验，例如EGFP实验，DRAQ5染色液要在激动剂和颉颃剂加入前的实验进程中（通常0.5～3h）加到培养基中，浓度控制在1μM。注：如果在DRAQ5染色前，细胞已经被别的荧光染料染色，注意上述操作过程要避光。 ⑤.染色细胞可直接进行相应分析，不需要清洗等别的操作。下表细胞数目及所需?DRAQ5体积及终浓度。
Product introduction 1. Specifications: 5mM 50 μL (suitable for 4 96-well experiments), 200 μL (suitable for 16 96-well experiments) 2. Parameters: MW= 412.49; Wavelength: Ex/Em= 646/697nm; Solvent: Water 3. Storage: 4 ℃ protected from light for 2 years Product description DRAQ5 is a far-infrared fluorescent living cell DNA dye with high affinity for double-stranded DNA. It is a dye that can penetrate the cell membrane and can mark live cells or fixed/dead cells. In flow cytometry, this dye can be used to distinguish between nucleated and non-nucleated cells. Because DRAQ5 can bind to DNA in a stoichiometric ratio, it can also be used to report nuclear DNA content, and is suitable for chromosome multiple and cell cycle analysis. In fluorescence microscopy analysis, it can be used as a nuclear counterstain. DRAQ5 has many applications and is highly compatible with programs widely used in existing instrument platforms, the main application areas are HCS, cell models, GFP, flow cytometry and fluorescence microscopes. The excitation wavelength range of DRAQ5 is 488～647nm. For imaging microscopy, it is recommended to use a 633 or 647nm light source for excitation. For flow cytometry, when the dye is excited at 488nm, the 685LP dichroic mirror and 710/50 channel can be used for detection; when excited at 633nm, the 660/20 channel can be used for detection. For cell cycle/DNA analysis applications, it is recommended to use a longer wavelength filter, such as 735LP dichroic mirror and 780/60 channel to optimize the CV value of G1 and G2/M peaks. Please make sure that your instrument can detect the dye. Due to the wide emission and excitation wavelength range of DRAQ, it is not recommended to combine DRAQ5 with other far-red fluorescent dyes that can be excited at 488 or 633 nm. Instructions (1) Description In the experiment, DRAQ5 is used as the last dye to stain, because DRAQ5 does not require the remaining washing steps after staining, so DRAQ5 can be directly added to the cell-containing medium for live cell staining (2) Operation ①. Prepare PBS buffer without sodium azide or a specific medium for specific cells. ②. Resuspend the cells in PBS or medium, and control the cell density to ≤ 4 × 105 cells/mL. For adherent cells and some tissues, roughly estimate the number of cells. ③. Add the appropriate volume of DRAQ5 staining solution of appropriate concentration according to Table 1. DRAQ5 staining solution can be directly added to the surface of tissues or adherent cells, or directly added to fresh medium ④. Mix gently, and incubate at room temperature for 5-30 minutes. Incubate at 37°C and shorten the time to 1-3min. For experiments with a long time span, such as the EGFP experiment, DRAQ5 staining solution should be added to the medium during the experiment before the agonist and antagonist are added (usually 0.5 to 3 hours), and the concentration is controlled at 1 μM. Note: If the cells have been stained with other fluorescent dyes before DRAQ5 staining, please keep away from light during the above operation. ⑤. The stained cells can be directly analyzed accordingly, without other operations such as washing. The following table shows the number of cells and the required volume and final concentration of DRAQ5.
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