计算溶液所需的质量、体积或浓度。
微丝染色液 (R250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
2-8°C储存,避光,室温。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
细胞骨架一般是指真核细胞胞质中纵横交错的纤维网，根据纤维直径、组成成分和组装结构的不同分为微管、微丝和中间纤维。观察细胞骨架的方法有电镜、组织化学、酶标记、免疫荧光等。微丝(microfilament)是由肌动蛋白构成的纤维，微丝在不同种类的细胞中与某些结合蛋白一起形成不同的亚细胞结构(如肌肉细丝、肠上皮微绒毛轴心、应力纤维等)。
微丝染色液 (R250法) 主要利用考马斯亮蓝R250显示由微丝构成的应力纤维，考马斯亮蓝R250可以对多种蛋白染色，并非特异染微丝，在该方法条件下，由于微管结构不稳定，有些类型的纤维太细，光学显微镜下无法辨认，因此能够看到的纤维主要是由微丝构成的应力纤维，直径约40nm，由于细胞对培养基质的附着和维持扁平铺展的形状，该纤维在体外培养的贴壁细胞中尤其发达。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：
| M1511591 | Component | 5×50mL | Storage |
| M1511591A | PBS 缓冲液 (10x) | 50mL | RT |
| M1511591B | TM 缓冲液 | 50mL | 2-8℃. Store in the dark. |
| M1511591C | M 缓冲液 (3x) | 50mL | 2-8℃. Store in the dark. |
| M1511591D | 微丝缓冲液 | 50mL | 2-8℃. Store in the dark. |
| M1511591E | 考马斯亮蓝 R250 染色液 | 50mL | RT |
操作步骤(仅供参考)：
（一）动物细胞微丝
1、取材：在盖玻片上细胞培养，生长密度达60～70%时细胞面朝上置于称量瓶中，用去离子水稀释PBS 缓冲液 (10x)至1×，用1×PBS 缓冲液清洗1min，重复1次。
2、抽提：弃PBS 缓冲液，加入2ml TM 缓冲液，盖上称量瓶盖子，37°C处理25～30min。
3、漂洗：弃TM 缓冲液，用去离子水稀释M 缓冲液(3×)至1×，用1×M 缓冲液清洗2min，重复2次。
4、固定：稍微晾干，加入2ml 微丝缓冲液固定细胞15～20min。
5、冲洗：弃微丝缓冲液，用1×PBS 缓冲液轻轻清洗2min，重复1次。
6、染色：弃PBS 缓冲液，把盖玻片立于吸水滤纸上吸去边缘水分，加入2ml 考马斯亮蓝 R250 染色液染色20～25min。
7、去离子水冲洗染液，滤纸吸干水分，晾干，镜检或树脂封片。
(二)植物细胞微丝
1、取材：用去离子水稀释PBS 缓冲液(10×)至1×，轻轻撕取约1 cm²洋葱鳞茎内皮，置于预先加入1×PBS 缓冲液的称量瓶中，孵育5～10min，使其下沉。
2、抽提：弃PBS 缓冲液，加入2ml TM 缓冲液，盖上称量瓶盖子，37°C处理30min。
3、漂洗：弃TM 缓冲液，用去离子水稀释M 缓冲液(3×)至1×，用1×M 缓冲液清洗3～5min，重复2次。
4、固定：稍微晾干，加入2ml 微丝缓冲液固定细胞20～25min。
5、冲洗：弃微丝缓冲液，用1×PBS 缓冲液清洗3～5min，重复2次。
6、染色：弃PBS 缓冲液，把盖玻片立于吸水滤纸上吸去边缘水分，加入2ml 考马斯亮蓝 R250 染色液染色20～25min。
7、去离子水冲洗染液，标本铺在载玻片上，加盖玻片，镜检。
染色结果：
光学显微镜下可见动物细胞轮廓，应力纤维呈深蓝色，形态长而直，常与细胞的长轴平行并贯穿细胞全长。洋葱表皮细胞轮廓清晰，微丝束呈深蓝色，高倍镜下，转到微调，可见细胞骨架的立体结构。
注意事项：
1、抽提、固定、染色应在加盖的称量瓶中进行，并且盖玻片的细胞面始终朝上。
2、沿称量瓶内壁缓慢加入各种试剂，避免直接滴到玻片或样本上；清洗细胞动作应轻柔，避免细胞脱落。3、抽提时间应自行摸索，时间过长易破坏细胞结构，时间过短易出现高背景。
4、应力纤维是一种动态结构，细胞充分贴壁时纤维挺拔、丰富；反之，细胞收缩变圆，应力纤维弯曲甚至部分解聚消失而显得稀少。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Cytoskeleton generally refers to the interlaced fiber network in the cytoplasm of eukaryotic cells. According to differences in fiber diameter, components and assembly structure, it can be divided into microtubules, microfilaments and intermediate filaments. Methods for observing the cytoskeleton include electron microscopy, histochemistry, enzyme labeling, immunofluorescence, etc. Microfilaments are fibers composed of actin. Together with certain binding proteins, microfilaments form different subcellular structures in various types of cells (such as muscle filaments, the core of intestinal epithelial microvilli, stress fibers, etc.).
Microfilament Staining Solution (R250 Method) mainly uses Coomassie Brilliant Blue R250 to reveal stress fibers composed of microfilaments. Coomassie Brilliant Blue R250 can stain a variety of proteins and is not specific to microfilaments. Under the conditions of this method, due to the instability of microtubule structure and the fact that some types of fibers are too thin to be recognized under an optical microscope, the fibers visible are mainly stress fibers composed of microfilaments with a diameter of approximately 40 nm. These fibers are particularly prominent in cultured adherent cells in vitro due to the cell’s attachment to the culture substrate and maintenance of a flat, spread morphology.
This reagent is for research use only and not intended for clinical diagnostic or other purposes.
Product Components and Storage Conditions:
| M1511591 | Component | 5×50mL | Storage |
| M1511591A | PBS Buffer(10x) | 50mL | RT |
| M1511591B | TM Buffer | 50mL | 2-8℃. Store in the dark. |
| M1511591C | M Buffer(3x) | 50mL | 2-8℃. Store in the dark. |
| M1511591D | Microfilament Fluid | 50mL | 2-8℃. Store in the dark. |
| M1511591E | R250 Stain | 50mL | RT |
Protocol (For Reference Only):
(I) Microfilaments in Animal Cells
1. Sample preparation: Culture cells on a coverslip. When the cell density reaches 60–70%, place the coverslip in a weighing bottle with the cell side up. Dilute PBS Buffer(10x) to 1× with deionized water, wash with 1× PBS Buffer for 1 min, and repeat once.
2. Extraction: Discard the PBS Buffer, add 2 mL of TM Buffer, cover the weighing bottle, and incubate at 37 °C for 25–30 min.
3. Rinsing: Discard the TM Buffer. Dilute 3× M Buffer to 1× with deionized water, wash with 1× M Buffer for 2 min, and repeat twice.
4. Fixation: Air-dry slightly, add 2 mL of Microfilament Fluid, and fix the cells for 15–20 min.
5. Washing: Discard the Microfilament Fluid, gently wash with 1× PBS Buffer for 2 min, and repeat once.
6. Staining: Discard the PBS Buffer. Stand the coverslip on absorbent filter paper to remove excess water from the edges, add 2 mL R250 stain, and stain for 20–25 min.
7. Rinse off the stain with deionized water, blot dry with filter paper, air-dry, and examine under a microscope or mount with resin.
(II) Microfilaments in Plant Cells
1. Sample preparation: Dilute PBS Buffer(10x) to 1× with deionized water. Gently tear off approximately 1 cm² of onion bulb epidermis, place it into a weighing bottle prefilled with 1× PBS Buffer, and incubate for 5–10 min until it sinks.
2. Extraction: Discard the PBS Buffer, add 2 mL of TM buffer, cover the weighing bottle, and incubate at 37 °C for 30 min.
3. Rinsing: Discard the TM Buffer. Dilute 3× M Buffer to 1× with deionized water, wash with 1× M Buffer for 3–5 min, and repeat twice.
4. Fixation: Air-dry slightly, add 2 mL of Microfilament Fluid, and fix the cells for 20–25 min.
5. Washing: Discard the Microfilament Fluid, wash with 1× PBS Buffer for 3–5 min, and repeat twice.
6. Staining: Discard the PBS Buffer. Stand the coverslip on absorbent filter paper to remove excess water from the edges, add 2 mL R250 stain, and stain for 20–25 min.
7. Rinse off the stain with deionized water, mount the specimen on a glass slide, cover with a coverslip, and examine under a microscope.
Staining Results:
Under an optical microscope, the outline of animal cells can be observed. Stress fibers appear dark blue, long and straight, often parallel to the long axis of the cell and extending through the entire length of the cell. The outline of onion epidermal cells is clear, and the microfilament bundles appear dark blue. Under high magnification, by adjusting the fine focus, the three‑dimensional structure of the cytoskeleton can be seen.
Precautions:
1. Extraction, fixation, and staining should be performed in a covered weighing bottle, with the cell‑bearing side of the coverslip always facing upward.
2. Add all reagents slowly along the inner wall of the weighing bottle to avoid direct contact with the coverslip or specimen. Handle cell washing gently to prevent cell detachment.
3. The extraction time should be optimized by the user: excessive time may damage cell structures, while insufficient time may result in high background.
4. Stress fibers are dynamic structures. When cells are fully adherent, the fibers are straight and abundant; otherwise, cells contract and round up, and stress fibers become bent, partially depolymerized, disappeared, or sparse.
5. For your safety and health, please wear a lab coat and disposable gloves during operation.
规格
别名
细胞骨架染色液 | 细胞骨架染色试剂盒
英文别名
Microfilament Stain
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
Microfilament Staining Solution (R250 Method)
储存条件
2-8°C储存,避光,室温
运输条件
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 |
|---|---|---|
 ZJ26F0433801 |
分析证书 | M1511591 |
技术文档和文章
溶液计算器
常见问题
什么是「生物染色剂」？
「生物染色剂」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
什么是「BioReagent」？
「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
本产品应如何储存？
请于2–8 °C、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。
本产品如何运输？
本产品采用湿冰冷藏运输。收货后请立即拆包，并按上述储存条件保存。
如何获取批次专用COA？
每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
需要帮助选择级别？
我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。
查看 BioReagent 级别指南 → 查看 Biological Stain 级别指南 → 查看 for Microscopy 级别指南 →产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）