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上海中乔新舟生物科技有限公司是美国Sciencell公司官网公布的一级代理商：
美国ScienCell研究实验室（www.sciencellonline.com）成立于1999年，公司总部位于美国加州的圣地亚哥。主要致力于实验室科研用原代细胞、原代细胞专用培养基、原代细胞无血清培养基、干细胞、干细胞培养基、干细胞无血清培养基的研究和开发，在全球拥有众多客户。在国内销售15年来，很多老师应用其产品发表了高质量的SCI文章，且有着极高的文献引用率。凭借着严格的质控和优秀的产品品质，深受广大科研工作者的信赖。
ScienCell研究实验室生产的原代细胞、原代细胞专用培养基都经过了严格的质量控制，细胞纯度可达98％。其中包括21种人体正常细胞系统，90多种不同细胞类型。大多数细胞在全球唯有ScienCell实验室能够成功分离，产品质量过硬。确保了实验结果的重复性和连贯性。
请登录Sciencell公司官方网站（www.zqxzbio.com或www.sciencellonline.com）以确保购买正规公司产品。
上皮细胞转染试剂盒 II说明：
货号：0973
Description
EpiFectagen II Cat. No. 0973, 250 transfections in 96-well plate .
Introduction
The delivery of foreign DNA into eukaryotic cells is one of the most common molecular biology techniques to study biological mechanisms. However, unlike transformed cell lines, the efficient transfection of primary cells can be a problem. EpiFectagen II is a cationic polymer-based transfection system specifically designed and optimized for efficient transfection of primary epithelial cells cultured in serum-free medium, such as Human Esophageal Epithelial Cells, Bronchial Epithelial Cells, Tracheal Epithelial Cells, Small Airway Epithelial Cells, Prostate Epithelial Cells, Corneal Epithelial Cells, Ovarian Surface Epithelial Cells and Mammary Epithelial Cells. Transfection with EpiFectagen II can be carried out in the presence of antibiotics. Instead of normal two-day transfection, an optimized one-day transfection procedure can be performed for time-saving and highly reproducible transfection.
Storage/Handling
Upon receipt, aliquot and store EpiFectagen II reagent A at -20°C, avoid repeated freezing/thawing cycles. Once thawed, store EpiFectagen II reagent A at 2-8°C and use in a month. EpiFectagen II reagent B can be kept at 2-8°C.
Quality Control
Each lot of EpiFectagen II is performance tested by transfecting Human Prostate Epithelial Cells (HPEpiCs, Cat. No. 4410, ScienCellT) with Promega® ?SV-bata-Galactosidase control vector. Gene expression is assayed by X-gal staining 24 hours post transfection. Typically, ~10% transfection efficiency can be achieved (Figure 1).
Procedures for Transfecting Adherent Cells in 96-well Plate*
- Preparation of cells
- On the day of transfection, coat 96-well plate with poly-l-lysine at 2 µg/cm2. Incubate at 37°C for 2-4 hours. Rinse the poly-l-lysine coated wells with sterile deionized H2O twice before seeding of cells. The pre-coating of poly-l-lysine ensures goodandeven epithelial cell adhesion.
- Select a flask of epithelial cells with 60-80% confluency, harvest and dilute cells in culture medium to give a final concentration of ~1.1×105 cells/ml.
- Transfection complex formation
- Prepare plasmid DNA in sterile deionized H2O to give a final concentration of 1 µg/µl. To achieve successful transfection, high quality DNA with OD260/OD 280 of 1.8 or greater is recommended.
- For each well, add 0.5 µl plasmid DNA, 12 µl sterile deionized H2O and 2 µl EpiFectagen II reagent B into a 1.5 ml sterile plastic tube. Vortex gently and spin down briefly. Then add 5.5 µl EpiFectagen II reagent A to make the total volume of the transfection mixture to be 20 µl, vortex for 5 seconds and spin down. Incubate at room temperature for 20-30 min.
- Incubation of cells with transfection mixture
- Plate 180 µl of cell suspension (~1.1×105 cells/ml) in each well to give ~2×104 cells per well.
- Add 20 µl of transfection mixture to each well. Mix by gently rocking the plate side-to-side.
- Culture the cells for ~ 24 hours under standard conditions. Or perform a medium change after 4-6 hours’ incubation with transfection mixtures, replace with 200 µl fresh culture medium, and culture for additional 16-18 hours. Generally longer incubation time with transfection mixture results in increased transfection efficiency and decreased cell viability.
- Harvest cells 24 hours post transfection and assay for gene expression.
* The amounts of cells and various transfection reagents mentioned in the instruction are recommended for performing transfection in 96-well plate. For transfection in larger size wells, the amounts of cells and transfection reagents (DNA, sterile deionized H2O and EpiFectagen II reagents A&B) should be scaled up according to the surface area of the wells (Table 1).
Table 1. Recommended quantities of epithelial cells and EpiFectagen II reagents per well.
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Culture Vessel
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Growth Area (cm2/well)
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# of cells
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1 µg/µl DNA stock (µl)
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Sterile DI H2O (µl)
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EpiFectagen II reagent B (µl)
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EpiFectagen II reagent A (µl)
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Culture Medium (µl)
|
|
96-well plate
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0.35
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20,000
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0.5
|
12
|
2
|
5.5
|
180
|
|
48-well plate
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0.8
|
45,000
|
1.1
|
27
|
4.6
|
12.6
|
411
|
|
24well plate
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2.0
|
115,000
|
2.9
|
69
|
11.6
|
31
|
1029
|
|
12-well plate
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4.0
|
230,000
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5.7
|
137
|
23
|
63
|
2057
|
|
6-well plate
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9.6
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550,000
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13.7
|
329
|
55
|
151
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4937
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如需要其他产品资料请发邮件至wwwfudan@163.com索取
Sciencell公司产品由上海中乔新舟生物科技有限公司优质供应
近几年的新药研发成功率在逐年下降，其根本原因之一就是传统的药物筛选系统是建立在只具有30-40%人类基因群的一系列细胞株上。这样一个有“缺陷型”药物筛选系统所产生的药物用于人体上就会出现许多致命的弱点和不完整性。新一代药物筛选系统是含有一系列近乎完整的人类基因群的原代细胞株。而利用这些原代细胞株所甄别和筛选出来的候选药物，其诊治人类疾病的成功几率将大大增加。这样不但大大节省了新药的开发成本，而且将极大地提高人类的健康质量。这些产品从根本上提高了全球生命医学研究、人类重要疾病药物研发、新药研发的成功率。所以上海中乔新舟生物科技有限公司致力于提供优质原代细胞产品和完善的售后服务。
美国ScienCell研究实验室（www.sciencellonline.com）成立于1999年，公司总部位于美国加州的圣地亚哥。主要致力于实验室科研用原代细胞、原代细胞专用培养基、原代细胞无血清培养基、干细胞、干细胞培养基、干细胞无血清培养基的研究和开发，在全球拥有众多客户。在国内销售15年来，很多老师应用其产品发表了高质量的SCI文章，凭借着严格的质控和优秀的产品品质，深受广大科研工作者的信赖。
ScienCell研究实验室生产的原代细胞、原代细胞专用培养基都经过了严格的质量控制，细胞纯度可达98％。其中包括21种人体正常细胞系统，90多种不同细胞类型。大多数细胞在全球唯有ScienCell实验室能够成功分离，产品质量过硬。 确保了实验结果的真实性、重复性和连贯性。
Sciencell公司部分原代细胞目录（如需要其他细胞资料请发邮件至 wwwfudan@163.com索取）：
中乔新舟www.zqxzbio.com专长于为生物医药领域的医疗机构、研究中心、企业、临床医生等提供课题设计、基金联合申请、实验技术服务、论文相关服务、采购外包等整体服务，目前已经成长为国内领先的转化医学外包品牌。
中乔新舟的PI团队已经发展到专职PI 20人，联席PI 312人，其中大多数拥有海外背景。截止到2009年3月，以PI或联席PI为第一作者发表SCI 论文1682 篇（总IF 值为5721.82，其中影响因子IF>5.0 的论文216 篇，IF>10.0论文32 篇）。
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