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Rat CD133 Human Cells Transfected Lysate (positive control) (denatured)
Catalog Number: orb174679
| Product Name | Rat CD133 Human Cells Transfected Lysate (positive control) (denatured) |
|---|---|
| Catalog Number | orb174679 |
| Hazard Information | Non-Toxic |
| Target | Rat CD133 |
| Form/Appearance | 50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue |
|---|---|
| Storage | Store at -80°C. Aliquot to avoid repeated freezing and thawing |
| Note | For research use only. |
| Biological Origin | Human Cells |
Human Cells transfected lysate in which Rat CD133/PROM1/Prominin 1 has been over-expressed. The whole cell lysate is provided in 1X Sample Buffer.
| Application Notes | Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined with Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol |
|---|
Material Safety Data Sheet
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如果经费充足的话可以找公司包装病毒,如武汉的普健可以提供各种载体的构建及细胞株构建的技术服务。
1.将外源基因插入慢病毒载体
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
先提取RNA,反转录成cDNA,然后根据目的基因设计PCR引物,通过半定量RT-PCR确定目的基因表达.
正因为检测的是mRNA,所以要先反转录成cDNA才能PCR.
最近因工作需要,欲购进一些生产用的细胞株和瞬转、稳转质粒,大家有什么推荐的品系和公司或网站,谢谢!!!
主要需求:
1.生产或生物制药用的,稳定细胞株如CHO-K1、GS和HEK293/HEK293T,以及与之对应的瞬转、稳转(重组整合)质粒(需详细图谱)。
2.原始或改造细胞株,以及对应质粒,改造细胞株请详述细节,原因,优缺点,安全性等。
3.希望大家积极推荐,实验用、生产用均可,原核、真核都行,网站或品系名称也行,只要大家觉得常用、稳定、安全、高产就行。
4.请有意向的同仁回帖或将详细资料发至我163邮箱:wangqiang23mars@163.com,万分感谢。

