DRAQ7是一款明亮的远红外亲脂性非膜透性蒽醌类DNA染料，其核心作用原理为通过特异性结合死细胞或透化细胞的细胞核DNA实现靶向标记，无法穿透完整活细胞的细胞膜，可以精准区分膜完整活细胞与膜受损死亡细胞。相较于死细胞标记染料PI与7-AAD，DRAQ7具备性能优势在于：激发波段覆盖广，发射光位于远红区域，不被紫外线激发、与PE及PE同系物无发射光谱重叠，可直接替代PI与7-AAD并规避其光谱交叉干扰的弊端；染色过程无需RNase处理以排除RNA干扰，也无需额外洗涤步骤，大幅简化实验流程；染料在细胞长期培养中无毒性，可保障实验样本的活性与后续研究的可行性。
DRAQ7凭借独特的标记特性与光谱优势，成为细胞凋亡、细胞活力及膜完整性研究的关键工具，因无长期毒性，DRAQ7还可作为药物筛选、细胞培养等实验的终点检测指标，用于评估不同处理条件下的细胞健康水平，为药物毒性、环境因子影响等研究提供可靠的细胞活力数据，其低干扰、高适配的特性有效突破了传统染料的应用局限，推动细胞死亡相关研究的精准化与高效化。仅限体外科研用途 不得用于临床诊断或治疗。
注：效能等同于Thermo Fisher DRAQ7™染料 (D15106)。
注：本文中DRAQ5、DRAQ7均是Biostatus Limited的商标及注册商标。
应用范围：
死细胞核酸检测、细胞凋亡、死细胞区分、多色共染分析等；
产品特点：
1、极速染色：室温最快10 min快速染色，大幅缩短实验时间；
2、免洗涤：染色后无需洗涤，方便使用，省时间省步骤；
3、方便使用：以易用的水溶性形式提供，按需稀释后即用，使用方便；
4、适合多色搭配：无紫外激发且发射光谱与PE无重叠，和PE搭配做凋亡检测时，无须设置单染管调节补偿；
5、超稳定：在室温下稳定，具有高度的热稳定性和光稳定性。
产品参数：
1、Ex：599/644 nm；
2、Em：678/697 nm；
3、光谱图：
产品组分：
| 组分 | 100 T | 1000 T | | Ready-to-use DRAQ7 Dead Cell DNA Dye（0.3 mM in Water） | 100 μL | 1 mL |
注：规格是按照DRAQ7母液1︰100稀释，工作浓度3 μM，稀释后96孔板检测单次用量100 μL来计算。 |
注意事项：
1、DRAQ7属于远红外荧光，肉眼不可见，需要CCD相机或激光共聚焦观察。
2、由于DRAQ7发射和激发波长范围很宽，不建议将Draq7与其他可被488 nm或633 nm激光激发的远红光荧光染料联用。
3、因叠氮化钠会影响DRAQ7染色效果，故实验中所用PBS等培养基或缓冲液均需要不含叠氮化钠。
4、荧光染料存在淬灭问题，请尽量注意避光，以减缓淬灭。
5、使用前请将产品瞬时离心，再进行后续操作。
6、本产品仅限于科研用途并且不得存放于普通住宅内。
7、为了您的安全和健康，请遵循您所在常规实验室安全规定。
使用说明：
一、 实验前准备
1. 试剂准备：
(1) 将试剂从储存条件 (如4 °C)取出，平衡至室温 (15-25 °C)。
(2) 准备细胞培养基、缓冲液 (如PBS)。
2. 对照设置
(1) 阴性对照：活细胞，新鲜完全培养基培养正常培养的细胞，作为标定“DRAQ7-活细胞”的荧光基线。
(2) 阳性对照：死细胞，55-60 ℃无菌蒸馏水孵育10 min (诱导完全细胞死亡)，标定“DRAQ7+死细胞”。
3. 仪器准备：
(1) 荧光显微镜：配备633 nm或647 nm光源的荧光显微镜。
(2) 流式细胞仪：可用633 nm波长激发的流式细胞仪。
二、 操作步骤
方案一：荧光显微镜检测 (适用于悬浮细胞)
1. 收集与洗涤细胞：
(1) 收集待测悬浮细胞悬液至离心管中。
(2) 1000 rpm，室温离心5 min，小心吸弃上清液。
2. 细胞重悬与计数：
(1) 使用PBS或细胞培养基重悬并细胞计数。
(2) 取5×10⁴-10⁵个细胞，1000 rpm低速离心5 min，去除PBS或细胞培养基。
3. 配制DRAQ7工作液：
将 DRAQ7按照1: 100的比例添加至细胞培养基中 (DRAQ7添加至培养基稀释比例在1: 15 至1: 200 之间)。
注：不同细胞种类所需DRAQ7的染色浓度可能不同，初期最好进行细胞染色的预实验工作，设置不同DRAQ7稀释倍数梯度确定最佳稀释比例。
4. 细胞染色：
(1) 将DRAQ7染色工作液添加至沉淀中，重悬细胞，室温 (20~25 ℃)孵育10 min；
(2) 孵育结束后，1000 rpm低速离心5 min，去除染色工作液；
(3) 加细胞培养基重悬细胞。
注：DRAQ7孵育时间建议在10 min上下摸索。
5. 荧光显微镜检测：
(1) 96孔板法：将100 μL细胞悬液加入96孔板中，静置片刻，待细胞自然沉降贴底后，置于荧光显微镜下观察。
(2) 载玻片法：取20-30 μL细胞悬液滴加于洁净载玻片上，轻轻盖上盖玻片，避免产生气泡，置于荧光显微镜下观察。
(3) 滤光片选择：Ex/Em: 645/666 nm。
方案二：荧光显微镜检测 (适用于贴壁细胞)
1. 细胞准备
提前一天在孔板中接种细胞，使得细胞汇合度达到70%-85%。
2. 配制DRAQ7工作液：
将DRAQ7按照1: 100的比例添加至细胞培养基中 (DRAQ7添加至培养基稀释比例在1: 15 至1: 200之间)。
注：不同细胞种类所需DRAQ7的染色浓度可能不同，初期最好进行细胞染色的预实验工作，设置不同DRAQ7稀释倍数梯度确定最佳稀释比例。
3. 细胞染色：
(1) 吸弃微孔板中的旧培养基；
(2) 稀释后的DRAQ7添加至细胞中 (染色工作液体积按建议添加：96 孔板加 100 μL/孔，48 孔板加150 μL/孔，24孔板加 250 μL/孔，12 孔板加 500 μL/孔，6 孔板加 1 mL/孔)，室温 (20~25 ℃)孵育10 min。
注：DRAQ7孵育时间建议在10 min上下摸索。
4. 荧光显微镜检测：
直接将培养板置于显微镜下检测，光片选择，Ex/Em: 645/666 nm。
方案三：流式细胞仪检测（适用于悬浮细胞和贴壁细胞）
1. 收集与制备单细胞悬液
(1) 悬浮细胞：同“方案一”中的“步骤1”和“步骤2”。
(2) 贴壁细胞：以6孔板培养的细胞为例。
a. 将细胞培养基吸出至适合的离心管内备用。贴壁细胞用少量PBS清洗一遍，清洗后的PBS吸出后同样转移到盛装培养基的离心管内，1000 rpm，室温离心5min。
b. 加入适量胰酶消化液 (覆盖细胞即可)，室温下置于显微镜下观察，待细胞变圆、间隙增大时，用移液器轻轻吹打，使细胞完全脱落。
注：胰酶使用量、消化时间、消化温度根据各自实验室的经验操作。
c. 关键：加入“a步骤”离心管内备用的细胞培养基，终止消化。
d. 将细胞悬液转移至离心管，1000 rpm，室温离心5 min，吸弃上清。
e. 收集a步骤与d步骤中的细胞。
f. 用PBS或细胞培养基重悬细胞并进行细胞计数。
g. 取5×10⁴-10⁵个细胞，1000 rpm，室温离心5 min，吸弃上清。
2. 配制DRAQ7染色工作液：
将DRAQ7按照1∶100的比例添加至细胞培养基中 (DRAQ7添加至培养基稀释比例在1: 15 至1: 200 之间)。
注：不同细胞种类所需DRAQ7的染色浓度可能不同，初期最好进行细胞染色的预实验工作，设置不同DRAQ7稀释倍数梯度确定最佳稀释比例。
3. 细胞染色：
使用1 mL DRAQ7染色工作液重悬细胞，室温 (20~25 ℃)孵育10 min。
注：DRAQ7孵育时间建议在10 min上下摸索。
4. 流式细胞仪检测：
选用633 nm波长激发，660 nm检测荧光信号。
三、 结果判读
1、定性分析 (显微镜)：
图 1. DRAQ7染料用于Hela细胞DNA染色的荧光显微成像
(1) 死细胞：细胞核区域出现明亮的远红外 (红色)荧光，信号集中且轮廓清晰，核区边界清晰；
(2) 活细胞：无核荧光，活细胞的细胞核区域无DRAQ7远红外 (红色)荧光信号 (因膜完整，染料无法进入)。
注：图片引自参考文献《Real-time cell viability assays using a new anthracycline derivative DRAQ7®》（doi: 10.1002/cyto.a.22228）
2、定性分析 (流式细胞仪)：
图 2. DRAQ7染料用于胶质瘤细胞的流式细胞图
(1) 第一步：根据阴性对照和阳性对照信号值确定“DRAQ7-活细胞群” 的基准和“DRAQ7⁺死细胞群”的基准；
(2) 第二步：FSC-A/SSC-A散点图，圈选细胞群，排除碎片与大聚集物；
(3) 活死细胞分群
a.FSC-A/DRAQ7散点图：辅助确认细胞大小与 DRAQ7 信号的关联，排除碎片干扰，x轴: FSC-A (线性轴，反映细胞大小)，y 轴: DRAQ7荧光强度 (双指数轴)
b.设定活/死阈值并圈门
定阈值：以“阴性对照 (活细胞)的DRAQ7信号”为基准，找到其荧光强度的第99百分位值 (即99%的活细胞信号低于该值)，此值即为 “活/死阈值”；
活细胞门 (DRAQ7⁻)：圈选“DRAQ7荧光强度低于阈值”的群体 (对应阴性对照的主要群体，信号弱)；
死细胞门 (DRAQ7⁺)：圈选“DRAQ7荧光强度高于阈值”的群体 (对应阳性对照的主要群体，信号强)。
注：图片引自参考文献《DRAQ7 as an Alternative to MTT Assay for Measuring Viability of Glioma Cells Treated With Polyphenols》（doi: 10.21873/anticanres.14553.）
常见疑问与解答：
1、问：DRAQ7储液是否含DMSO？
答：该产品为水溶液形式，不含DMSO。
2、问：染色时间在37 ℃和室温下为什么有差异？
答：37 ℃时细胞的活性和分子运动相对室温更活跃，DRAQ7与细胞核结合的速度更快，所以可能需要更短的染色时间。
3、问：在使用DRAQ5和DRAQ7的时候，这两者有什么区别，该如何选择？
答：对活细胞进行 DNA 分析、长期监测细胞状态或进行组织深层成像时选DRAQ5；评估细胞活力、检测凋亡或区分样本中的死活细胞群体时选DRAQ7。两者的区别如下：
| 对比 | DRAQ5 | DRAQ7 | | 细胞膜渗透性 | 可透过 | 不可透过 | | 染色对象 | 活细胞和固定细胞的所有细胞核 | 仅死亡细胞、凋亡细胞或透化细胞的细胞核 | | 结合方式 | 与DNA化学计量结合，荧光强度反映DNA含量 | 特异性结合双链DNA，染色快速 | | 适用场景 | 活细胞DNA含量分析、细胞周期研究、长期活细胞成像实验、深层组织/肿瘤组织成像 | 细胞活力评估、细胞凋亡检测、药物/化合物体外毒性测试、区分死活细胞的流式细胞术实验 |
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DRAQ7 is a bright far-red lipophilic non-membrane-permeable anthraquinone DNA dye. Its core mechanism of action is to achieve targeted labeling by specifically binding to nuclear DNA of dead or permeabilized cells. It cannot penetrate the cell membrane of intact live cells, enabling accurate differentiation between membrane-intact live cells and membrane-damaged dead cells. Compared with PI and 7-AAD, the classic dead cell labeling dyes, DRAQ7 has the following performance advantages: it features a broad excitation wavelength range with emission in the far-red region, is not excited by ultraviolet light, and has no emission spectral overlap with PE and its analogs, which allows it to directly replace PI and 7-AAD and avoid the drawbacks of their spectral cross-interference; the staining process requires no RNase treatment to eliminate RNA interference or additional washing steps, greatly simplifying the experimental workflow; the dye is non-toxic in long-term cell culture, ensuring the viability of experimental samples and the feasibility of subsequent research.
With its unique labeling characteristics and spectral advantages, DRAQ7 has become an essential tool for studies on cell apoptosis, cell viability and membrane integrity. Due to its lack of long-term toxicity, DRAQ7 can also serve as an endpoint detection indicator for experiments such as drug screening and cell culture, used to evaluate cell health under different treatment conditions and provide reliable cell viability data for research on drug toxicity, the effects of environmental factors and other related fields. Its low-interference and high-compatibility properties effectively break through the application limitations of traditional dyes and promote the precision and efficiency of cell death-related research. For in vitro research use only. Not for clinical diagnosis or treatment.
Note: Equipotent to Thermo Fisher DRAQ7™ Dye (D15106).Note: DRAQ5 and DRAQ7 in this document are trademarks and registered trademarks of Biostatus Limited.
Applications
Nucleic acid detection of dead cells, cell apoptosis analysis, dead cell discrimination, multicolor co-staining analysis, etc.
Product Features
1. Rapid staining: Fast staining in as little as 10 minutes at room temperature, significantly shortening experimental time;
2. No washing required: No post-staining washing steps, offering ease of use and saving time and operational procedures;
3. User-friendly: Supplied as an easy-to-use water-soluble formulation, ready for use after dilution on demand;
4. Suitable for multicolor combination: No UV excitation and no emission spectral overlap with PE; no single-stain tube is needed for compensation adjustment when combined with PE for apoptosis detection;
5. Ultra-stable: Stable at room temperature with high thermal and photostability.
Product Specifications
1. Ex: 599/644 nm;
2. Em: 678/697 nm;
3. Spectra:
Product Components
| Component | 100 T | 1000 T | | Ready-to-use DRAQ7 Dead Cell DNA Dye（0.3 mM in Water） | 100 μL | 1 mL |
Note: The specifications are calculated based on a 1:100 dilution of DRAQ7 stock solution (working concentration: 3 μM) and a single usage volume of 100 μL for 96-well plate detection after dilution. |
Precautions
1. DRAQ7 is a far-red fluorescent dye, invisible to the naked eye, and requires observation with a CCD camera or laser confocal microscope.
2. Due to the broad excitation and emission wavelength range of DRAQ7, it is not recommended to use Draq7 in combination with other far-red fluorescent dyes excitable by 488 nm or 633 nm laser.
3. Sodium azide interferes with the staining effect of DRAQ7; therefore, all media or buffers such as PBS used in experiments must be sodium azide-free.
4. Fluorescent dyes are prone to quenching; please avoid light exposure as much as possible to slow down quenching.
5. Centrifuge the product briefly before use before performing subsequent operations.
6. This product is for research use only and must not be stored in ordinary residential premises.
7. For your safety and health, follow the general laboratory safety regulations of your institution.
Instructions for Use
I. Pre-Experiment Preparation
1. Reagent Preparation
(1) Take the reagent out of storage conditions (e.g., 4 °C) and equilibrate to room temperature (15-25 °C).
(2) Prepare cell culture medium and buffer (e.g., PBS).
2. Control Setting
(1) Negative control: Live cells, normally cultured in fresh complete medium, used to calibrate the fluorescence baseline of "DRAQ7⁻ live cells".
(2) Positive control: Dead cells, incubated in sterile distilled water at 55-60 °C for 10 min (to induce complete cell death), used to calibrate "DRAQ7⁺ dead cells".
3. Instrument Preparation
(1) Fluorescent microscope: Equipped with a 633 nm or 647 nm light source.
(2) Flow cytometer: Capable of excitation at 633 nm wavelength.
II. Operational Procedures
Protocol 1: Fluorescent Microscopy Detection (for Suspension Cells)
1. Cell Collection and Washing
(1) Collect the suspension cell suspension to be tested into a centrifuge tube.
(2) Centrifuge at 1000 rpm for 5 min at room temperature, and carefully aspirate and discard the supernatant.
2. Cell Resuspension and Counting
(1) Resuspend cells with PBS or cell culture medium and perform cell counting.
(2) Take 5×10⁴-10⁵ cells, centrifuge at low speed (1000 rpm) for 5 min, and remove PBS or cell culture medium.
3. Preparation of DRAQ7 Working Solution
Add DRAQ7 to the cell culture medium at a ratio of 1:100 (the dilution ratio of DRAQ7 in the medium can range from 1:15 to 1:200).
Note: The optimal staining concentration of DRAQ7 may vary with cell types. It is recommended to perform a pre-experiment for cell staining initially and set up a gradient of different DRAQ7 dilution ratios to determine the best dilution ratio.
4. Cell Staining
(1) Add the DRAQ7 staining working solution to the cell pellet, resuspend the cells, and incubate at room temperature (20~25 °C) for 10 min;
(2) After incubation, centrifuge at low speed (1000 rpm) for 5 min and remove the staining working solution;
(3) Resuspend cells with cell culture medium.
Note: It is recommended to optimize the DRAQ7 incubation time around 10 minutes.
5. Fluorescent Microscopy Detection
(1) 96-well plate method: Add 100 μL of cell suspension to a 96-well plate, let it stand for a while until the cells settle to the bottom naturally, and observe under a fluorescent microscope.
(2) Slide method: Drop 20-30 μL of cell suspension onto a clean glass slide, gently cover with a coverslip to avoid air bubbles, and observe under a fluorescent microscope.
(3) Filter selection: Ex/Em: 645/666 nm.
Protocol 2: Fluorescent Microscopy Detection (for Adherent Cells)
1. Cell Preparation
Seed cells in a well plate one day in advance to achieve a cell confluency of 70%-85%.
2. Preparation of DRAQ7 Working Solution
Add DRAQ7 to the cell culture medium at a ratio of 1:100 (the dilution ratio of DRAQ7 in the medium can range from 1:15 to 1:200).
Note: The optimal staining concentration of DRAQ7 may vary with cell types. It is recommended to perform a pre-experiment for cell staining initially and set up a gradient of different DRAQ7 dilution ratios to determine the best dilution ratio.
3. Cell Staining
(1) Aspirate and discard the old medium in the microplate;
(2) Add the diluted DRAQ7 to the cells (recommended volume of staining working solution: 100 μL/well for 96-well plate, 150 μL/well for 48-well plate, 250 μL/well for 24-well plate, 500 μL/well for 12-well plate, 1 mL/well for 6-well plate), and incubate at room temperature (20~25 °C) for 10 min.
Note: It is recommended to optimize the DRAQ7 incubation time around 10 minutes.
4. Fluorescent Microscopy Detection
Place the culture plate directly under the microscope for detection with filter setting of Ex/Em: 645/666 nm.
Protocol 3: Flow Cytometry Detection (for Both Suspension and Adherent Cells)
1. Cell Collection and Single-Cell Suspension Preparation
(1) Suspension cells: Follow Step 1 and Step 2 in Protocol 1.
(2) Adherent cells: Take cells cultured in a 6-well plate as an example.
a. Aspirate the cell culture medium into a suitable centrifuge tube for later use. Rinse the adherent cells with a small amount of PBS once, transfer the rinsed PBS to the centrifuge tube containing the medium, and centrifuge at 1000 rpm for 5 min at room temperature.
b. Add an appropriate amount of trypsin digestion solution (just covering the cells), place at room temperature and observe under a microscope; once the cells become rounded and the intercellular spaces increase, gently pipette to detach the cells completely.
Note: The dosage of trypsin, digestion time and digestion temperature shall be determined according to the laboratory's established protocols.
c. Key step: Add the cell culture medium reserved in step a to terminate digestion.
d. Transfer the cell suspension to a centrifuge tube, centrifuge at 1000 rpm for 5 min at room temperature, and aspirate and discard the supernatant.
e. Collect the cells from step a and step d.
f. Resuspend cells with PBS or cell culture medium and perform cell counting.
g. Take 5×10⁴-10⁵ cells, centrifuge at 1000 rpm for 5 min at room temperature, and aspirate and discard the supernatant.
2. Preparation of DRAQ7 Staining Working Solution
Add DRAQ7 to the cell culture medium at a ratio of 1:100 (the dilution ratio of DRAQ7 in the medium can range from 1:15 to 1:200).
Note: The optimal staining concentration of DRAQ7 may vary with cell types. It is recommended to perform a pre-experiment for cell staining initially and set up a gradient of different DRAQ7 dilution ratios to determine the best dilution ratio.
3. Cell Staining
Resuspend the cells with 1 mL of DRAQ7 staining working solution and incubate at room temperature (20~25 °C) for 10 min.
Note: It is recommended to optimize the DRAQ7 incubation time around 10 minutes.
4. Flow Cytometry Detection
Select 633 nm wavelength for excitation and detect the fluorescent signal at 660 nm.
III. Result Interpretation
1. Qualitative Analysis (Microscopy)
Figure 1. Fluorescent micrographs of Hela cell DNA stained with DRAQ7 dye
(1) Dead cells: Bright far-red (red) fluorescence appears in the nuclear region with concentrated, well-defined signals and clear nuclear boundaries;
(2) Live cells: No nuclear fluorescence, and no DRAQ7 far-red (red) fluorescent signal is detected in the nuclear region of live cells (the dye cannot enter due to intact cell membranes).
Note: The image is cited from the reference Real-time cell viability assays using a new anthracycline derivative DRAQ7® (doi: 10.1002/cyto.a.22228).
2. Qualitative Analysis (Flow Cytometry)
Figure 2. Flow cytometry plot of glioma cells stained with DRAQ7 dye
(1) Step 1: Determine the baseline of "DRAQ7⁻ live cell population" and "DRAQ7⁺ dead cell population" according to the signal values of negative and positive controls;
(2) Step 2: On the FSC-A/SSC-A scatter plot, gate the cell population to exclude debris and large aggregates;
(3) Live/dead cell gating
a. FSC-A/DRAQ7 scatter plot: Assist in confirming the correlation between cell size and DRAQ7 signal to exclude debris interference (X-axis: FSC-A (linear axis, reflecting cell size); Y-axis: DRAQ7 fluorescence intensity (bi-exponential axis)).
b. Set the live/dead threshold and gate the populationsThreshold setting: Based on the DRAQ7 signal of the negative control (live cells), find the 99th percentile of its fluorescence intensity (i.e., 99% of live cell signals are below this value), which is defined as the "live/dead threshold";
Live cell gate (DRAQ7⁻): Gate the population with DRAQ7 fluorescence intensity below the threshold (corresponding to the main population of the negative control with weak signals);
Dead cell gate (DRAQ7⁺): Gate the population with DRAQ7 fluorescence intensity above the threshold (corresponding to the main population of the positive control with strong signals).
Note: The image is cited from the reference DRAQ7 as an Alternative to MTT Assay for Measuring Viability of Glioma Cells Treated With Polyphenols (doi: 10.21873/anticanres.14553.).
Frequently Asked Questions (FAQs)
1. Q: Does the DRAQ7 stock solution contain DMSO?
A: This product is a water-based formulation and contains no DMSO.
2. Q: Why is there a difference in staining time between 37 °C and room temperature?
A: Cell activity and molecular movement are more vigorous at 37 °C than at room temperature, and DRAQ7 binds to the cell nucleus at a faster rate, so a shorter staining time may be required.
3. Q: What are the differences between DRAQ5 and DRAQ7 in use, and how to choose between them?
A: Choose DRAQ5 for DNA analysis of live cells, long-term monitoring of cell status, or deep tissue imaging; choose DRAQ7 for evaluating cell viability, detecting apoptosis, or distinguishing live/dead cell populations in samples. The differences between the two are as follows:
| Comparison | DRAQ5 | DRAQ7 | | Cell membrane permeability | Permeable | Non-permeable | | Staining target | All cell nuclei of live and fixed cells | Only cell nuclei of dead, apoptotic or permeabilized cells | | Binding mode | Stoichiometric binding to DNA, fluorescence intensity reflects DNA content | Specific binding to double-stranded DNA with rapid staining | | Applicable scenarios | Live cell DNA content analysis, cell cycle research, long-term live cell imaging experiments, deep tissue/tumor tissue imaging | Cell viability assessment, cell apoptosis detection, in vitro toxicity testing of drugs/compounds, flow cytometry experiments for distinguishing live/dead cells |
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