Lipofectine
1. Simplest transfection protocol – ready to use, just mix with DNA and vortex;
2. Superior transfection efficiency;
3. Effective for both adherent and suspension cells;
4. Minimal cytotoxicity;
5. Change of cell culture media after transfection is recommended but not required;
6. Highly reproducible results;
7. Product stable at 4°C and at -20°C.
Protocol for transfection of adherent cells:
1. For each microgram of DNA to be transfected, add 50 ul of Lipofectine;
2. Flick, rock, pipet up and down, or vortex the mix for 5 seconds;
3. Incubate the mix at room temperature for 15 minutes;
4. Add the mix to media and cells.
Note: Change of media 12 to 24 hour post transfection may improve cell growth condition, and is recommended. With Lipofectine, change of media is not absolutely required.
Protocol for transfection of suspension cells:
Lipofectine has proven to be an effective reagent to transfect suspension cells. The ideal cell density at time of transfection is 1 x 106 per ml. Therefore, the day before transfection, suspension CHO cells are seeded at 0.5 x 106 per ml in media such as ProCHO (Lonza) or CD-CHO (Invitrogen) without serum. The amount of DNA used in transfection should be 3 µg per ml of cells, which yields optimal cell growth and target protein expression. Transfection mix can be prepared exactly as described in the previous section. The transfection mix is then added to the suspension cells. Media replacement is recommended 12 to 24 hour post transfection, and data has shown that media replacement can increase cell growth rate after transfection for suspension. For protein or antibody production, cell media can be harvested 4-7 days post transfection.
Note: Each cell line and plasmid is different. The protocols and recommendations are for CHO cells. Users should optimize the condition for their cell lines and plasmids.
Biowit Technologies Ltd. is a high-tech company specialized in providing R&D services and related products in the biological field. As a technology-driven and customer-focused company, BioWit provides a broad and integrated service and product portfolio including molecular biology, virus packaging (e.g., adenovirus, AAV, lentivirus, retrovirus and baculovirus), protein expression, cell biology, stem cells and transgenic models. One of the milestones achieved by Biowit is its recent sucess in the development of a series of serum-containing media as well as serum-free or animal -free media for stem cell culture, which are tailored for clinical applications. The core technology team members have engaged in research and development for many years in world-famous academic institutions or prestigious drug companies. Biowit has established collaboration with international prestigious research institutions. The worldwide collaboration enables Biowit to timely renew its biotechnologies (http://www.biowit.com)
百恩维生物科技有限公司（BioWit Technologies），是一家专业从事生物技术研发并提供相关技术服务和产品的高新技术企业。公司目前提供的技术服务和产品主要包括分子生物学技术服务、腺病毒（Adenovirus)、慢病毒(Lentivirus)、腺相关病毒（AAV）、杆状病毒（Baculovirus)等包装技术服务、蛋白质表达与纯化技术服务（包括原核细菌、杆状病毒-昆虫细胞、哺乳动物细胞等）、高效转染试剂（HET Kit)、细胞因子(如hLif、mLif、aFGF、bFGF、EGF、TGFα、TGFβ等）、干细胞培养基(如胎胚干细胞培养基、间充质干细胞培养基等）、干细胞的分离和存储以及个体特异性诱导型多能干细胞的制备和存储, 其中干细胞无血清或无动物源培养是专门为临床打造和设计的快速扩增细胞的培养基同时又能很好地维持其干细胞的潜能。公司的长期目标是推出人类重大疑难疾病的干细胞治疗及基因治疗临床方案和预防措施，重点研究神经疾病、视网膜疾病、白血病、血友病、艾滋病和肝炎的干细胞治疗和基因治疗方案，及提供高效的预防动物或人的传染病、流行病的疫苗(http://www.biowit.com)。
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