计算溶液所需的质量、体积或浓度。
小鼠外周血淋巴细胞分离液
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为什么选择此级别
BioReagent,无菌过滤 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。常规运输 。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
外周血单个核细胞 (peripheral blood mononuclear cell, PBMC)是指外周血中具有单个核的细胞，包含淋巴细胞、单核细胞、树突状细胞和其它少量细胞 (造血干细胞等)。PBMC是免疫学功能研究中最常用的细胞模型，如细胞增殖、细胞毒性、细胞因子分泌等。
小鼠外周血淋巴细胞分离液主要成分是聚蔗糖和泛影酸钠，是一种用于分离小鼠外周血淋巴细胞的无菌、低内毒素水平的密度梯度分离液。适用于从小鼠血液及组织匀浆中分离所需细胞。其分离原理是根据血细胞的密度差异 (分离小鼠单个核细胞时比重为1.080；分离大鼠单个核细胞时比重为1.084-1.087，红细胞和粒细胞的密度为1.092g/mL左右，血小板为1.030-1.035g/mL之间)，通过离心使一定密度的细胞按相应密度梯度分布，从而将淋巴细胞从小鼠外周血或血中分离出来。
产品性能指标：
外观：本产品为乳光或微乳光的注射水溶液
密度：1.081±0.001 g/mL
渗透压：280-340 mOsmol/kg
无菌：0.22μm 滤膜过滤
该产品仅用于科研领域，不适用于临床诊断或其他用途。
操作步骤 (仅供参考)：
1、稀释血液样本: 1份的等渗溶液 (如PBS缓冲液)加1-2份的血液进行稀释，即等渗溶液: 抗凝血=1: 1-2。
2、为满足客户实际使用需求，根据不同血液样本量推荐具体实验操作方法如下：
情况A：血液样本量小于2mL时，实验方法如下：
1、取一支15mL离心管，加入4mL分离液。
2、用吸管小心吸取血液样本加于分离液之液面上，450-500g，离心20-30min（注：根据血液样本量确定离心条件，血液样本量越多，离心力越大，离心时间越长，具体离心条件需客户自行摸索，以达到最佳分离效果）。
3、离心后，此时离心管中由上至下分为四层。第一层为血浆层。第二层为环状乳白色淋巴细胞层。第三层为透明分离液层。第四层为红细胞层。如下图示：
4、用吸管小心吸取第二层环状乳白色淋巴细胞层到另一15mL离心管中，向所得离心管中加入10mL等渗溶液，混匀细胞。
5、250g，离心10min。
6、弃上清。
7、用吸管以5mL等渗溶液重悬所得细胞。
8、250g，离心10min。
9、重复6、7、8，弃上清后以1mL后续实验所需相应液体重悬细胞。
情况B：血液样本量为2-4mL时，实验方法如下：
1、取一支15mL离心管，加入5mL分离液。
2、用吸管小心吸取血液样本加于分离液之液面上，450-650g，离心20-30min（注: 根据血液样本量确定离心条件，血液样本量越多，离心力越大，离心时间越长，具体离心条件需客户自行摸索，以达到最佳分离效果，但最大离心力最好不超过1200g）。
3、离心后，此时离心管中由上至下分为四层。第一层为血浆层。第二层为环状乳白色淋巴细胞层。第三层为透明分离液层。第四层为红细胞层。
4、用吸管小心吸取第二层环状乳白色淋巴细胞层到另一15mL离心管中，向所得离心管中加入10mL等渗溶液），混匀细胞。
5、250g，离心10min。
6、弃上清。
7、用吸管以5mL等渗溶液重悬所得细胞。
8、250g，离心10min。
9、重复6、7、8，弃上清后以1mL后续实验所需相应液体重悬细胞。
注意事项：
1、开封前颠倒混匀，本分离液为无菌产品，易感染细菌，需无菌条件操作，无菌条件下启封后置于4℃保存，有效期6个月。
2、分离液使用时应始终保持室温 (18℃-25℃)，如室内温度较低，可将分离液预热。4℃或者是温度较低的条件下离心，可能会导致白膜层中红细胞污染加重。
3、血液样本最好为新鲜抗凝血 (采血2h以内)，为保持淋巴细胞的活性，应避免冷冻和冷藏。
4、稀释血液或洗涤细胞，不可使用含Ca、Mg离子的缓冲液及培养液，其成分会导致血细胞凝集，大大降低细胞得率及纯度。
5、部分塑料制品 (如聚苯乙烯）因其带有的静电作用，可能会导致细胞挂壁，影响分离效果。
6、血液样本的粘稠度或者是温度差异，可能会影响分离效果，可以调节离心转数和离心时间，摸索最佳的分离条件。
7、吸取过多的淋巴细胞层及分离液层会导致分离液交界处的粒细胞被吸出从而使混杂的粒细胞数量增加；吸取过多的血浆层可能会导致淋巴细胞中血浆蛋白及血小板污染。
8、如果要进一步对分离的细胞进行培养，那在收集血液和分离过程中，注意无菌操作，避免微生物污染。
9、不同动物血液在不同比重分离液中的细胞离散系数及细胞带电不同，用户在制定分离液时应提供所需分离液的比重、动物种属及被分离细胞的名称。
10、本产品对光敏感，应该室温避光储存。
11、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Peripheral blood mononuclear cells (PBMCs) refer to mononuclear cells in peripheral blood, including lymphocytes, monocytes, dendritic cells, and other minor cell populations (e.g., hematopoietic stem cells). PBMCs are the most commonly used cellular model for immunological function studies, such as cell proliferation, cytotoxicity, and cytokine secretion assays.
Mouse peripheral blood lymphocyte separation medium is primarily composed of Ficoll and sodium diatrizoate. It is a sterile density gradient separation medium with low endotoxin levels, specifically designed for isolating mouse peripheral blood lymphocytes. It is suitable for separating target cells from mouse blood and tissue homogenates. The separation principle is based on the density differences among blood cells (the optimal density is 1.080 for isolating mouse mononuclear cells; 1.084-1.087 for rat mononuclear cells; the density of red blood cells and granulocytes is approximately 1.092 g/mL; and the density of platelets ranges from 1.030 to 1.035 g/mL). Through centrifugation, cells of specific densities distribute according to the corresponding density gradient, thereby separating lymphocytes from mouse peripheral blood or tissues.
Product Performance Specifications
Appearance: Opalescent or slightly opalescent aqueous injection solution
Density: 1.081±0.001 g/mL
Osmolality: 280-340 mOsmol/kg
Sterility: Filtered through a 0.22 μm membrane
This product is for research use only and is not intended for clinical diagnosis or any other purposes.
Operating Procedures (For Reference Only)
1. Dilute the blood sample: Mix 1 volume of isotonic solution (e.g., PBS buffer) with 1-2 volumes of blood, resulting in a ratio of isotonic solution: anticoagulated blood=1:1-2.
2. To meet practical experimental needs, the recommended operational protocols based on different blood sample volumes are as follows:
Scenario A: Blood sample volume<2 mL
1. Take a 15 mL centrifuge tube and add 4 mL of separation medium.
2. Carefully pipette the diluted blood sample and layer it gently onto the surface of the separation medium. Centrifuge at 450-500×g for 20-30 min. (Note: Centrifugation parameters depend on the blood sample volume. A larger sample volume requires higher centrifugal force and longer centrifugation time. Users should optimize these parameters to achieve the best separation efficiency.)
3. After centrifugation, the contents in the centrifuge tube are separated into four distinct layers from top to bottom:
Layer 1: Plasma layer
Layer 2: Opaque milky white lymphocyte layer
Layer 3: Transparent separation medium layer
Layer 4: Red blood cell layer
(As illustrated in the figure below)
4. Carefully aspirate the second layer (opaque milky white lymphocyte layer) into a new 15 mL centrifuge tube. Add 10 mL of isotonic solution to the tube and resuspend the cells thoroughly.
5. Centrifuge at 250×g for 10 min.
6. Discard the supernatant.
7. Resuspend the collected cells with 5 mL of isotonic solution using a pipette.
8. Centrifuge at 250×g for 10 min.
9. Repeat steps 6, 7, and 8. After discarding the final supernatant, resuspend the cells in 1 mL of the appropriate medium required for subsequent experiments.
Scenario B: Blood sample volume=2-4 mL
1. Take a 15 mL centrifuge tube and add 5 mL of separation medium.
2. Carefully pipette the diluted blood sample and layer it gently onto the surface of the separation medium. Centrifuge at 450-650×g for 20-30 min. (Note: Centrifugation parameters depend on the blood sample volume. A larger sample volume requires higher centrifugal force and longer centrifugation time. Users should optimize these parameters to achieve the best separation efficiency. The maximum centrifugal force is recommended not to exceed 1200×g.)
3. After centrifugation, the contents in the centrifuge tube are separated into four distinct layers from top to bottom:
Layer 1: Plasma layer
Layer 2: Opaque milky white lymphocyte layer
Layer 3: Transparent separation medium layer
Layer 4: Red blood cell layer
4. Carefully aspirate the second layer (opaque milky white lymphocyte layer) into a new 15 mL centrifuge tube. Add 10 mL of isotonic solution to the tube and resuspend the cells thoroughly.
5. Centrifuge at 250×g for 10 min.
6. Discard the supernatant.
7. Resuspend the collected cells with 5 mL of isotonic solution using a pipette.
8. Centrifuge at 250×g for 10 min.
9. Repeat steps 6, 7, and 8. After discarding the final supernatant, resuspend the cells in 1 mL of the appropriate medium required for subsequent experiments.
Precautions
1. Invert and mix well before opening. This separation medium is a sterile product and is susceptible to bacterial contamination; therefore, all operations must be performed under sterile conditions. After opening under sterile conditions, store the product at 4°C, with a validity period of 6 months.
2. The separation medium should be maintained at room temperature (18°C-25°C) during use. If the ambient temperature is too low, pre-warm the medium appropriately. Centrifugation at 4°C or lower temperatures may increase red blood cell contamination in the buffy coat layer.
3. The blood sample should preferably be fresh anticoagulated blood (collected within 2 hours). To maintain lymphocyte viability, avoid freezing and refrigeration of the sample.
4. For blood dilution or cell washing, do not use buffers or culture media containing Ca²⁺ or Mg²⁺-these ions can cause blood cell agglutination, which significantly reduces cell yield and purity.
5. Some plastic materials (e.g., polystyrene) may cause cell adhesion due to electrostatic effects, thereby affecting separation efficiency.
6. Variations in blood sample viscosity or temperature may impact separation results. Adjust centrifugation speed and duration to optimize separation conditions.
7. Aspirating an excessive amount of the lymphocyte layer or separation medium layer may lead to the inclusion of granulocytes at the interface, increasing granulocyte contamination. Aspirating an excessive amount of the plasma layer may result in contamination of lymphocytes with plasma proteins and platelets.
8. If further culture of the isolated cells is required, ensure strict sterile operation during blood collection and separation processes to avoid microbial contamination.
9. Different animal blood samples exhibit varying cell dispersion coefficients and cell charge properties in separation media of different densities. When preparing custom separation media, users should specify the required medium density, animal species, and the name of the target cells to be isolated.
10. This product is light-sensitive and should be stored at room temperature in the dark.
11. For your safety and health, wear a lab coat and disposable gloves during operation.
规格
规格或纯度
BioReagent,无菌过滤
稳定性与储存
Store at room temperature long term (24 months). Store in the dark.
英文名称
Mouse Peripheral Blood Lymphocyte Separation Medium
储存条件
避光,室温
运输条件
常规运输
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
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0333491 |
分析证书 | M1509600 |
技术文档和文章
溶液计算器
常见问题
什么是「无菌过滤」？
「无菌过滤」表示该物料经过加工和检测，适用于特种工业加工。规格侧重于影响该应用性能的杂质——例如，电子应用的金属含量、电池应用的粒度、光学应用的折射率，或对水分敏感工艺的水分含量。
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本产品应如何储存？
请于室温、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？
本产品按常温标准条件运输，无需温控包装。
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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。
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。
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