计算溶液所需的质量、体积或浓度。
细胞自噬染色检测试剂盒 (MDC法)
有货
为什么选择此级别
BioReagent,生物染色剂,用于显微镜 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
避光,室温,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
自噬（autophagy）是细胞受到刺激后吞噬自身的细胞质或细胞器，最终将吞噬物在溶酶体内降解的过程，自噬体（autophagosome）为双层膜包被的圆形或椭圆形结构，内含细胞质、长寿蛋白质和异种蛋白聚集物、损伤或多余细胞器如线粒体、粗面内质网和微体、病毒和细菌等。自噬是一种进化上保守的降解过程，通过溶酶体途径将长寿命蛋白、细胞器及其他细胞质等组分进行降解，自噬通路的激活对多种细胞功能都是必需的，包括饥饿状态下的存活、细胞内组分清除、发育及免疫等过程。丹酰尸胺（Dansylcadaverine，MDC），是一种嗜酸性的荧光色素，通常被用于检测自噬体形成的特异性标记染色剂，其检测激发滤光片波长355~380nmnm，阻断滤光片波长512~530nm。细胞自噬染色检测试剂盒（MDC法）又称为MDC染色液，适用于培养细胞的荧光自噬染色观察，又可用FACS检测细胞自噬发生的比例，可与EB合用双染。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
产品组分及储存条件：
| C1511655 | Component | 100T | Storage |
| C1511655A | MDC Stain(10x) | 1mL | -20℃. Store in the dark. |
| C1511655B | Stain Buffer | 50mL | RT |
| C1511655C | Wash Buffer | 100mL | RT |
自备材料：
1、低速离心机、1.5ml离心管、载玻片、盖玻片、96孔板、荧光显微镜、流式细胞仪
2、培养液、蒸馏水、磷酸盐缓冲液
操作步骤（仅供参考）：
（一）玻片法
1、收集细胞，用300～500μl的Wash Buffer 清洗细胞1次，800～1000rpm离心5min，弃上清，加入适量的Stain Buffer重悬细胞，计数并调节细胞浓度至10⁶/ml。
2、取90μl细胞悬液至新的1.5ml离心管中，加入10μl的 MD CStain(10×)，轻轻混匀。
3、37°C或室温避光染色15～60min。
4、800～1000rpm离心5min，弃上清，收集细胞，用300～500μl的Wash Buffer清洗细胞2次，800～1000rpm离心5min，弃上清。
5、加入100μl的Wash Buffer重悬细胞，滴加于载玻片上并加盖玻片。
6、荧光显微镜观察计数并拍照，也可用流式细胞仪检测药物干预组MDC阳性细胞百分比。
（二）96孔板法
1、轻轻吸除96孔板中的培养液，各孔加入10μl MDC Stain(10×）和90μl的Stain Buffer，37°C 5% CO₂ 条件下避光孵育15～60min。
2、各孔加入100μl Wash Buffer清洗2～3次，荧光显微镜或流式细胞仪检测观察。
（三）MDC与EB双染法
1、收集细胞，用300～500μl的Wash Buffer 清洗细胞1次，800～1000rpm离心5min，弃上清，加入适量的Stain Buffer重悬细胞，计数并调节细胞浓度至10⁶/ml。
2、取90μl细胞悬液至新的1.5ml离心管中，加入10μl MDC Stain(10×）和0.2uM EB染色液，轻轻混匀，滴加于载玻片上，室温避光染色15～60min，加盖玻片。
3、荧光显微镜或流式细胞仪检测观察。
染色结果：
荧光染色结果：药物干预组细胞呈现了更高的荧光强度和更多的亮绿色MDC标记颗粒；
FACS检测结果：药物干预组MDC阳性细胞百分比高于对照组。
注意事项：
1、MDC Stain和EB对人体有一定害处，请小心操作。
2、收集细胞前可在培养液中加入不同浓度的药物培养一定时间，诱导细胞自噬。
3、操作过程中应注意减少试剂暴露于强光下的时间。
4、Wash Buffer可用PBS代替。
5、试剂开封后请尽快使用，以防影响后续实验效果。
6、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Autophagy is a process in which cells engulf their own cytoplasm or organelles in response to stimulation, and ultimately degrade the engulfed materials within lysosomes. Autophagosomes are spherical or elliptical structures surrounded by a double membrane, containing cytoplasm, long‑lived proteins and aggregates of abnormal proteins, damaged or redundant organelles such as mitochondria, rough endoplasmic reticulum and microbodies, as well as viruses and bacteria. Autophagy is an evolutionarily conserved degradation process that degrades long‑lived proteins, organelles, and other cytoplasmic components via the lysosomal pathway. Activation of the autophagic pathway is essential for diverse cellular functions, including survival under starvation, clearance of intracellular components, development, and immune responses. Dansylcadaverine (MDC) is an acid‑responsive fluorescent dye commonly used as a specific marker for detecting autophagosome formation. Its excitation filter wavelength is 355–380 nm, and its barrier filter wavelength is 512–530 nm. The Cellular Autophagy Staining Detection Kit (MDC Method), also known as MDC Staining Solution, is suitable for fluorescent staining and observation of autophagy in cultured cells. It can also be used to detect the proportion of autophagic cells by FACS, and can be used for double staining in combination with EB. This reagent is for research use only and is not intended for clinical diagnosis or other applications.
Product Components and Storage Conditions:
| C1511655 | Component | 100T | Storage |
| C1511655A | MDC Stain(10x) | 1mL | -20℃. Store in the dark. |
| C1511655B | Stain Buffer | 50mL | RT |
| C1511655C | Wash Buffer | 100mL | RT |
Materials Required:
1. Low-speed centrifuge, 1.5 mL centrifuge tubes, glass slides, cover slips, 96-well plates, fluorescence microscope, flow cytometer
2. Cell culture medium, distilled water, phosphate-buffered saline (PBS)
Protocol (For Reference Only)
(I) Slide Method
1. Collect cells. Wash cells once with 300–500 μL Wash Buffer, centrifuge at 800–1000 rpm for 5 min, and discard the supernatant. Resuspend cells in an appropriate volume of Stain Buffer, count cells, and adjust the concentration to 1×10⁶ cells/mL.
2. Transfer 90 μL of cell suspension to a new 1.5 mL centrifuge tube, add 10 μL MDC Stain (10×), and mix gently.
3. Stain at 37 °C or room temperature in the dark for 15–60 min.
4. Centrifuge at 800–1000 rpm for 5 min, discard the supernatant, and collect cells. Wash cells twice with 300–500 μL Wash Buffer, centrifuge at 800–1000 rpm for 5 min, and discard the supernatant.
5. Resuspend cells in 100 μL Wash Buffer, mount on a glass slide, and cover with a cover slip.
6. Observe, count, and capture images under a fluorescence microscope. The percentage of MDC-positive cells in the drug-treated group can also be detected by flow cytometry.
(II) 96-Well Plate Method
1. Gently remove the culture medium from the 96-well plate. Add 10 μL MDC Stain (10×) and 90 μL Stain Buffer to each well. Incubate at 37 °C in 5% CO₂ in the dark for 15–60 min.
2. Add 100 μL Wash Buffer to each well and wash 2–3 times. Detect and observe with a fluorescence microscope or flow cytometer.
(III) MDC & EB Double Staining
1. Collect cells. Wash cells once with 300–500 μL Wash Buffer, centrifuge at 800–1000 rpm for 5 min, and discard the supernatant. Resuspend cells in an appropriate volume of Stain Buffer, count cells, and adjust the concentration to 1×10⁶ cells/mL.
2. Transfer 90 μL of cell suspension to a new 1.5 mL centrifuge tube, add 10 μL MDC Stain (10×) and 0.2 μM EB Stain, mix gently. Mount on a glass slide, stain at room temperature in the dark for 15–60 min, and cover with a cover slip.
3. Detect and observe with a fluorescence microscope or flow cytometer.
Staining Results:
1. Fluorescence microscopy: The drug-treated group showed stronger fluorescence intensity and more bright green MDC-labeled puncta.
2. FACS analysis: The percentage of MDC-positive cells in the drug-treated group was higher than that in the control group.
Precautions:
1. MDC Stain and EB are harmful to humans; handle with care.
2. Before cell collection, cells can be cultured with different concentrations of drugs for a certain period to induce autophagy.
3. Minimize exposure of reagents to strong light during operation.
4. Wash Buffer can be replaced with PBS.
5. Use the reagents promptly after opening to avoid affecting experimental results.
6. For your safety and health, wear a lab coat and disposable gloves during operation.
规格
别名
细胞自噬染色试剂盒 | MDC染色液
英文别名
Cell Autophagy Staining Kit (MDC Method) | Autophagy/Cytotoxicity Dual Stain Kit (MDC Method)
规格或纯度
BioReagent,生物染色剂,用于显微镜
稳定性与储存
各组分在相应的储存条件下保质期1年。
英文名称
Autophagy Staining and Detection Kit (MDC Method)
储存条件
避光,室温,-20°C储存
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0434149 |
分析证书 | C1511655 |
技术文档和文章
溶液计算器
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