货号: | LM-T1702 |
数量: | 大量 |
供应商: | 法国Biosera |
规格: | 500 ml |
LM-T1702 | Lymphosep, Lymphozyte Separation Media | ||||
State : Liquid | Storage : + 20 °C | Life : 24 months | Size : | 100 ml 500 ml | |
Composition - Safety datasheet - Technical datasheet |
Lymphosep , Lymphozyte Separation Media
CAT N : LM-T1702
Theoretical pH : 7.0 ± 0.5
Osmolarity : 300 ± 20 mOsm/l
Colour : colourless, clear solution Storage conditions : Room temperature
Shelf life : 24 months Density: 1.077 ± 0.001
Sterility tests :
- bacteria in aerobic and anaerobic conditions
- fungi and yeast
Endotoxin : < 10 EU/ml (<1ng/ml)
Composition : Displayed on website and in catalogue; also available on request
Recommended use :
Use aseptic techniques when handling this medium.Product is provided for in vitro laboratory use only, and not for drug, human or veterinary use.
Applications :
Lymphosep is designed for the simple, rapid isolation of lymphocytes from whole blood that has been diluted and treated with anti-coagulant or defibrinating agent.For best results use blood drawn less than 2 hours before. Do not use blood more than 24 hours from when it was drawn.
Uses :
1) Thoroughly mix the Lymphosep by inverting the bottle gently.
2) Aseptically transfer 3 ml of Lymphosep to a 15 ml centrifuge tube.
3) Mix 2 ml of defibrinated or hepranized blood with 2 ml of physiological saline (PBS w/o Ca w/o Mg) or balanced salt solution
4) Carefully layer the diluted blood over 3 ml of Lymphosep (room temperature) in a 15 ml centrifuge, creating a sharp blood-Lymphosep interphase. DO NOT MIX! The quality of the separation is dependent upon a sharp interphase between the lymphocytes and the solution.
5) Centrifuge the tube at 400G at room temperature for 15 to 30 minutes. Centrifugation should sediment erythrocytes and polynuclear leukocytes and band mononuclear lymphocytes above the Lymphosep.
6) Aspirate the top layer of clear plasma to within 2-3 mm above the lymphocyte layer.
7) Aspirate the lymphocyte layer plus about half of the Lymphosep layer below it and transfer it to a centrifuge tube. Add an equal volume of buffered balanced salt solution to the lymphocyte layer in the centrifuge tube and centrifuge for 10 minutes at room temperature (18°C to 25°C) at a speed sufficient to sediment the cells without damage i.e., 160-260 g. Washing the cells removes Lymphosep and reduces the percentage of patelets.
8) Wash the cells again with buffered balanced salt solution and resuspend in the appropriate medium for your applications.
中文翻译,仅供参考:
备注:该产品相当于sigma公司的Histopaque®-1077产品,也相当于GE公司的Ficoll-Paque PREMIUM,欢迎选购。
人淋巴细胞分离液
货号: LM-T1702
理论 pH : 7.0 ± 0.5
渗透压: 295 ± 10 mOsm/l
颜色: 无色, 透明液体
贮存条件: 室温
保质期: 24个月
无菌检测:
- 细菌分别在有氧和无氧环境中检测
- 真菌和酵母检测
内毒素含量: < 10 EU/ml (<1ng/ml)
成分: 网站和目录上的说明; 您也可以跟我们获取
推荐使用:
请在使用该分离液时进行无菌操作。
该产品只能用于科研,不能用于药物,人兽使用。
应用:
淋巴细胞分离液能够简单,快速的从全血中分离淋巴细胞,并且是经过稀释以及用抗凝剂或除去血液中的纤维蛋白试剂处理的。
要想得到最好的结果,血液放置不要超过2个小时,不要使用放置超过24小时的血液进行实验。
使用方法:
1) 温和颠倒瓶子,充分混匀淋巴细胞分离液。
2) 无菌转移3 ml 淋巴细胞分离液到一支15 ml离心管里.
3) 将2 ml 去纤维蛋白和肝素的血液与2 ml 生理盐水 (PBS w/o Ca w/o Mg) 或者平衡盐溶液相混合。
4) 在15 ml离心中,小心的将稀释的血液加入3 ml 淋巴细胞分离液的液面上(室温),在血液和淋巴细胞分离液中间形成一个明显的分层。不要混合!因为分离的质量取决于淋巴细胞和溶液之间的那个小薄层界面。
5) 室温下400g离心15-30分钟,离心可以沉淀红细胞和多核白细胞同时可以在淋巴细胞分离液上形成一层单核淋巴细胞。
6) 吸出淋巴细胞上方2-3mm的透明血浆。
7) 吸取淋巴细胞层以及它下面一半的淋巴细胞分离液,然后转移到另外的一个离心管中。加入等体积的平衡盐缓冲液至淋巴细胞离心管中,室温(18°C to 25°C)离心10分钟,离心速度设定在既不损伤细胞又能沉淀细胞即可,例如160 - 260 g(去除淋巴细胞分离液和降低血小板的百分比)。
8) 用平衡盐缓冲液清洗细胞,用适当的培养基重悬细胞。