计算溶液所需的质量、体积或浓度。
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。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 130 篇同行评审文献引用。
概述
储存缓冲液：磷酸盐溶液
伊文思蓝 (Evans Blue)又称偶氮蓝，分子式C₃₄H₂₄N₆Na₄O₁₄S₄，分子量为960.81，CAS号为314-13-6。伊文思蓝属于一种常用的偶氮染料制剂，因其分子量大小与血浆白蛋白相近，而且在血液中与血浆白蛋白有很高的亲和力，因此在神经科学研究中常被用于示踪观察血脑屏障 (BBB)的完整性，也用于细胞染色区分活细胞、死细胞，亦可测定血容量。伊文思蓝作临床药物用于测定血浆和血容量，也可作动脉插管的定位。正常情况下血浆白蛋白无法透过血脑屏障，所以染色后如果神经系统血脑屏障完整，与血浆白蛋白结合的伊文思蓝无法使其着色，相反如果神经系统血脑屏障被破坏，伊文思蓝就可以进入神经系统并使其着色，在荧光波长470nm、540nm处有强峰，680 nm处有弱峰，可以使用化学透析法和比色法进行检测。
伊文思蓝与台盼蓝都是细胞活性染料，常用于检测细胞膜的完整性和细胞是否存活；活细胞不会被染成蓝色，而死细胞会被染成淡蓝色，伊文思蓝染色后通过显微镜下直接计数或显微镜下拍照后计数，就可以对细胞存活率进行比较精确的定量，其中0.5%为最常用的浓度；活细胞因有外排功能而无法被伊文思蓝染色，因此可以通过此方法在显微镜下区分死细胞与活细胞，但无法区分死亡与坏死。伊文思蓝染色液 (0.1%)主要由伊文思蓝、磷酸盐溶液等组成。该试剂仅用于科研领域，不用于临床诊断或其他用途。
自备材料：
1、注射器、组织匀浆器
2、PBS、三氯乙酸或丙酮
操作步骤(仅供参考)：
(一)血脑屏障通透性
1、取处理后的实验动物 (以小鼠为例)，经尾静脉或股静脉按照 2-3ml/kg的比例注射Evans Blue Staining Solution (0.1%)数秒至1分钟内，小鼠眼睛、皮肤出现蓝色，0.5-1h后处死小鼠，取目的脑组织。
2、脑组织置于1.5ml离心管中，加入1ml PBS，迅速用组织匀浆器将脑组织制成匀浆, 1000g离心15min。
3、取上清，加入等量三氯乙酸，4℃孵育18-24h；该步骤亦可采用如下操作：取上清，按上清：丙酮=3：7比例加入丙酮，室温孵育24h。
4、1000g离心20-30 min或2000g离心15min。
5、取上述溶液1-2ml，用分光光度计测620 nm处吸光值 (OD值)，同时测定已知不同梯度的标准伊文思蓝的OD值，绘制标准曲线，根据标准曲线计算出待测待测样品的伊文思蓝含量。
(二)活细胞染色
1、取100μl重悬细胞到常规1.5ml或0.5ml离心管内，加入100μl Evans Blue Staining Solution (0.1%)轻轻混匀，染色3min (染色时间可适当延长，但不宜超过10min)。
2、吸取少量经过染色后的细胞，用血细胞计数板计数。通常如果要比较精确地进行定量，每个细胞样品至少数500个细胞，数出蓝色细胞和细胞总数。计算公式如下：
细胞存活率＝(细胞总数－蓝色细胞数)/细胞总数×100%
(三)种子染色
1、用刀片做横切和沿种胚中央准确纵切，加入染色液染色3-5min。
2、蒸馏水中浸泡20-60min，视脱色程度而定。
(四)石蜡切片IFA衬染
1、常规烤片、脱蜡。
2、第二抗体含一定浓度的伊文斯蓝 (0.1%)衬染30 min。
3、缓冲甘油封片观察。
(五)植物整体叶片活细胞检测
1、取胁迫处理后的叶片用纯水洗净，干燥，浸泡在 0.25% (W/V)的伊文斯蓝中24h。
2、清水清洗表面蓝色染液，吸去表面水分放入煮沸的无水乙醇：甘油 (9：1)中30min，除去叶绿素至叶片泛白，拍照记录蓝色叶斑的叶片。
3、将叶片剪碎，放入4mL SDS溶液 (1%)中萃取3d，以沸水煮15~20min后被伊文斯蓝全部染上蓝色的叶片作为对照，提取液在600nm下测定吸光值。
注意事项：
1、Evans Blue Staining Solution (0.1%)对人体有轻微毒性，请小心防护。
2、细胞染色时，注意凋亡小体偶尔也有拒染现象。
3、血脑屏障通透性实验中，Evans Blue Staining Solution (0.1%)注射量应根据不同动物以及动物的重量调整。
4、最好采用低温冷冻离心机进行离心。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Evans Blue, also known as azo blue, has a molecular formula of C₃₄H₂₄N₆Na₄O₁₄S₄, a molecular weight of 960.81, and a CAS number of 314-13-6. As a commonly used azo dye preparation, Evans Blue has a molecular weight similar to that of plasma albumin and high affinity with plasma albumin in the blood. Therefore, it is often used in neuroscience research to trace and observe the integrity of the blood-brain barrier (BBB). It is also used for cell staining to distinguish between live and dead cells, and for blood volume determination. Clinically, Evans Blue is used as a drug to measure plasma and blood volume, and also for the positioning of arterial cannulation. Under normal circumstances, plasma albumin cannot penetrate the blood-brain barrier. Thus, if the BBB in the nervous system remains intact after staining, Evans Blue bound to plasma albumin will not stain the nervous system. On the contrary, if the BBB is damaged, Evans Blue can enter the nervous system and stain it. Evans Blue shows strong absorption peaks at fluorescence wavelengths of 470 nm and 540 nm, and a weak peak at 680 nm, and can be detected by chemical dialysis and colorimetry.
Both Evans Blue and trypan blue are cell viability dyes, commonly used to detect the integrity of cell membranes and cell viability. Live cells will not be stained blue, while dead cells will be stained light blue. After Evans Blue staining, the cell survival rate can be quantified relatively accurately by direct counting under a microscope or counting after microscopic photography. The most commonly used concentration is 0.5%. Live cells cannot be stained by Evans Blue due to their efflux function, so this method can be used to distinguish dead cells from live cells under a microscope, but cannot distinguish between apoptosis and necrosis. Evans Blue Staining Solution (0.1%) is mainly composed of Evans Blue and phosphate solution. This reagent is for research use only and not for clinical diagnosis or other purposes.
Materials to Be Prepared by Users
1. Syringes, tissue homogenizers
2. PBS (Phosphate-Buffered Saline), trichloroacetic acid, or acetone
Operating Procedures (for Reference Only)
(1) Blood-Brain Barrier Permeability Test
1. Take the treated experimental animals (mice as an example), and inject Evans Blue Staining Solution (0.1%) via the tail vein or femoral vein at a dose of 2-3 ml/kg. Within a few seconds to 1 minute, the mice’s eyes and skin will turn blue. Sacrifice the mice 0.5-1 hour later and collect the target brain tissue.
2. Place the brain tissue in a 1.5 ml centrifuge tube, add 1 ml of PBS, and quickly homogenize the brain tissue using a tissue homogenizer. Centrifuge at 1000×g for 15 minutes.
3. Collect the supernatant, add an equal volume of trichloroacetic acid, and incubate at 4°C for 18-24 hours. Alternatively, the following operation can be adopted: collect the supernatant, add acetone at a ratio of supernatant:acetone = 3:7, and incubate at room temperature for 24 hours.
4. Centrifuge at 1000×g for 20-30 minutes or at 2000×g for 15 minutes.
5. Take 1-2 ml of the above solution, measure the absorbance (OD value) at 620 nm using a spectrophotometer. Meanwhile, determine the OD values of standard Evans Blue solutions with different gradients, draw a standard curve, and calculate the Evans Blue content in the sample to be tested based on the standard curve.
(2) Live Cell Staining
1. Take 100 μl of resuspended cells into a conventional 1.5 ml or 0.5 ml centrifuge tube, add 100 μl of Evans Blue Staining Solution (0.1%), mix gently, and stain for 3 minutes (the staining time can be appropriately extended but should not exceed 10 minutes).
2. Aspirate a small amount of stained cells and count them using a hemocytometer. For relatively accurate quantification, at least 500 cells should be counted for each cell sample, and the number of blue cells and the total number of cells should be recorded. The calculation formula is as follows:
Cell Survival Rate = [(Total Number of Cells - Number of Blue Cells) / Total Number of Cells] × 100%
(3) Seed Staining
1. Use a blade to make cross-sections and accurate longitudinal sections along the center of the seed embryo, then immerse the sections in the staining solution for 3-5 minutes.
2. Soak the stained sections in distilled water for 20-60 minutes, depending on the degree of decolorization.
(4) IFA Counterstaining for Paraffin Sections
1. Perform routine slide baking and dewaxing.
2. Incubate with the secondary antibody containing Evans Blue at a specific concentration (0.1%) for counterstaining, lasting 30 minutes.
3. Mount the sections with buffered glycerol and observe under a microscope.
(5) Viable Cell Detection in Intact Plant Leaves
1. Take the leaves treated with stress, wash them thoroughly with pure water, dry them, and soak them in 0.25% (W/V) Evans Blue solution for 24 hours.
2. Rinse the surface of the leaves with clean water to remove the blue staining solution, blot the surface moisture, and immerse the leaves in boiling absolute ethanol:glycerol (9:1) solution for 30 minutes. Continue until chlorophyll is removed and the leaves turn pale, then take photos to record the leaves with blue spots.
3. Cut the leaves into pieces, place them in 4 mL of 1% SDS solution for extraction over 3 days. Use leaves fully stained blue by Evans Blue (after boiling in water for 15-20 minutes) as the control. Measure the absorbance of the extract at 600 nm.
Precautions
1. Evans Blue Staining Solution (0.1%) has slight toxicity to the human body; please take careful protective measures.
2. During cell staining, note that apoptotic bodies may occasionally exhibit dye exclusion (i.e., not be stained).
3. In the blood-brain barrier permeability test, the injection volume of Evans Blue Staining Solution (0.1%) should be adjusted according to the type and weight of the experimental animals.
4. It is recommended to use a low-temperature refrigerated centrifuge for centrifugation.
5. For your safety and health, please wear a lab coat and disposable gloves during operation.
规格
别名
伊文思蓝染色液；偶氮蓝染色液
英文别名
Evans Blue Staining Solution
规格或纯度
BioReagent,生物染色剂,用于显微镜,0.1%
稳定性与储存
Store at 2-8℃ long term (12 months). Store in the dark.
英文名称
Evans Blue Staining Solution (0.1%)
储存条件
2-8°C储存,避光
运输条件
低温运输
产品线
生物产品
名称和识别符
EC号
206-242-5
分子类型
生物试剂/缓冲液
Isomeric SMILES
CC1=C(C=CC(=C1)C2=CC(=C(C=C2)N=NC3=C(C4=C(C=C3)C(=CC(=C4N)S(=O)(=O)[O-])S(=O)(=O)[O-])O)C)N=NC5=C(C6=C(C=C5)C(=CC(=C6N)S(=O)(=O)[O-])S(=O)(=O)[O-])O.[Na+].[Na+].[Na+].[Na+]
分子量
960.81
Beilstein号
16(2)259
Reaxy-Rn
7110511
Reaxys-RN link address
https://www.reaxys.com/reaxys/secured/hopinto.do?context=S&query=IDE.XRN=7110511&ln=
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
安全和危险性（GHS）
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
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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 |
|---|---|---|
 ZJ26F0737405 |
分析证书 | E1507841 |
| ZJ26F0737144 |
分析证书 | E1507841 |
技术文档和文章
此产品的引用文献
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。
本产品的 CAS 号、分子式和分子量是多少？
本产品的 CAS 号为 314-13-6。
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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 →相关产品
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