等级
for molecular biology
质量水平
形式
dried film
用途
kit sufficient for 75-200 transfections
存货情况
available only in USA, Canada and EU
技术
transfection: suitable
储存温度
2-8°C
相关类别
一般描述
应用
- C6胶质瘤细胞（人类）
- 皮质神经元细胞（大鼠原代细胞）
- 背根神经节(DRG)细胞（大鼠）
- NT2神经细胞（人前体细胞）
- NT神经元（人分化细胞）
- 室下区 (SVZ)细胞（小鼠）
- 白质细胞（小鼠）
特点和优势
- 针对原代神经细胞、神经胶质细胞以及培养的神经细胞系进行了优化
- 具有超低的细胞毒性，不会造成神经细胞退化或树突戒断
- 可有效地递送原代神经细胞、神经胶质细胞以及培养的神经细胞系
- 相对于其他方法，使用更为轻松、快捷
- 兼容含血清和不含血清的转染实验
组分
1.5 mL缓冲液 H9036
7.5 mL DNA稀释液D1941
注意
原理
法律信息
储存分类代码
10 - Combustible liquids
闪点(°F)
Not applicable
闪点(°C)
Not applicable
分析证书(COA)
输入产品批号来搜索 分析证书(COA) 。批号可以在产品标签上"批“ （Lot或Batch）字后找到。
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示例
其它示例：
705578-5MG-PW
PL860-CGA/SHF-1EA
MMYOMAG-74K-13
1000309185
输入内容 1.000309185)
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Aldrich 产品
如果您查询到的批号为 TO09019TO 等，请输入去除前两位字母的批号：09019TO。
如果您查询到的批号含有填充代码（例如05427ES-021），请输入去除填充代码-021的批号：05427ES。
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未找到您寻找的产品？
部分情况下，可能未在线提供COA。如果搜索不到COA，可在线索取。
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
Why do I see a precipitate in my cell culture after lipid-based transfection?
The precipitate is likely excess lipid or EDTA and will probablly not affect transfection efficiency. If your DNA plasmid is suspended in TE, be sure the concentration of EDTA is <0.3 mM, or suspend the DNA in sterile molecular biology grade water instead.
Is low cell passage number an important consideration for transfection?
Yes, we recommend cells are at a low passage when being used for any application, including transfection. The reason why depends on what type of cells they are. Primary cells will undergo a finite number of divisions, and as they get closer to senesence they divide more slowly - both affecting their ability to take up DNA (transient transfection), and minimizing their abillity to incorporate the DNA into the genome (stable selection).Cultured common cell lines are often immortalized, and generally continue to aquire mutations, leading to a heterogenous population that may perform differently from cells of lower passage number - leading to results that are not reproducible.
Is the size of the plasmid an important consideration for transfection?
The size of the plasmid should be considered when selecting a transfection reagent with the best efficiency. In general, larger sized plasmids should easily transfect with readily available transfection reagents, as along as the plasmid DNA is of high purity.
Is optimizing the transfection protocol important?
For many common cell lines, transfection reagent efficiency is very high and the protocols will not require any optimization. For hard-to-transfect cells or those ultimately expressing a toxic protein, the protocol should be optimized for best transfection efficiency. Taking time to optimize will give you more transfected cells with each procedure, which can mean more protein expressed and results that are reproducible.
How do I choose a transfection reagent?
There are many guides that help you select a transfection reagent. In general, consider:The type of cell(s) you will transfectThe type of nucleic acid or protein you will introduce to the cellThe composition of your cell culture mediumThe need for stable or transient transfectionThe equipment you have availableThe other factors important to you - cost, protocol flexibility, ease of use, etc.
What quality does the DNA need to be in order to use it for transfection?
The DNA needs to be good quality or it may cause the cells to lyse and/or they won't transfect efficiently. Plasmid DNA prepared with a column-based DNA purification kit is suitable for transfections. Sigma's GenElute™ Minprep, Midiprep and Maxiprep kits work well for DNA plasmid purification. After preparing the DNA, confirm the OD A260:A280 ratio is greater than 1.6 for use in plasmid transfections.
What is transfection efficiency?
Transfection efficiency is a measure of how many cells take up the DNA during the transfection process. Many transfection reagents can achieve a transfection efficiency of >90% in common cell lines. Other cell lines are hard to transfect, and require special reagents and/or techniques to achieve even a small population of transfected cells.
How can I determine the efficiency of my transfection?
Calculating transfection efficiency is very useful when optimizing transfection protocols. Transfection efficiency can be performed using a GFP-expressing plasmid. After transfection, cells are stained with propidium iodide and counted. The propidium iodide provides a count of the total cells in the population, and the GFP-expressing cells provide a count of the number of cells transfected. The transfection efficiency (%) can then be calculated by:(# GFP-expressing cells / total cell #) * 100
How can I increase the efficiency of my transfection?
Transfection efficiency is affected by many different things, including plasmid size and purity, media components present, transfection reagent selected, amount of DNA and transfection reagent used, cell density, etc. Optimizing the protocol with respect to these concerns will allow you to achieve a higher transfection efficiency. For many cell lines and transfection reagents, optimized protocols are already available.
Can I transfect cells plated at low density?
For most transfections, cells should be >70% confluency the day of transfection, and growing in mid-log phase. Some transfection reagents are now designed to work with cells at low density, when required.
Can antibiotics be present in the medium during transfection?
We recommend that no antibiotics are present during transfection. The process of transfection can make the cells somewhat more porous to allow for efficient DNA entry. During this time, antibiotics will also enter the cells more easily and the cells may show increased cell death. Wait until about 24 hours after transfection to resume the use of preventative antibiotics and/or start the use of selective antibiotics.
Why are neurons difficult to transfect?
Neurons are both non-dividing cells, and sensitive to toxicity from the transfection reagent used. Sigma's Neuroporter™ Transfection Reagent is an optimized transfection reagent with a ready-to-use protocol that is helping overcome these limitations.
What is the difference between stable and transient transfection?
When the DNA enters the nucleus of the cell, the plasmid is replicated by the cell machinery (transient transfection). During this time, RNA is transcribed and protein translated until the plasmid DNA is lost after a few cell divisions. This expression of the plasmid DNA, mRNA, and protein is transient (temporary).In some cases, the plasmid DNA is integrated into the host cell genome. This is usually accompanied by forced expression using a selection antibiotic and sometimes a cloning step (to be sure all cells have the same integration site). Once the DNA is stable, the cell line can be frozen and used to express protein for many years. Clones may even be screened for those expressing the highest amount of protein.
My question is not addressed here, how can I contact Technical Service for assistance?
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商品
Transfection is the introduction of DNA, RNA, or proteins into eukaryotic cells and is used in research to study and modulate gene expression. Thus, transfection techniques and protocols serve as an analytical tool that facilitates the characterization of genetic functions, protein synthesis, cell growth and development.
转染是将DNA、RNA或蛋白质引入真核细胞的过程，用于研究和调节基因表达。因此，转染技术和实验方案作为分析工具，有助于表征遗传功能、蛋白质合成、细胞生长和发育。
This brief webinar provides an overview of what transfection is and the methods that are used to introduce DNA or RNA into eukaryotic cells.
相关内容
Browse our convenient transfection reagent selection guide to match the best reagent for your specific cell line and application needs.
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