跳轉至內容
Merck
全部照片(3)

Key Documents

D1163

Sigma-Aldrich

DOTAP硫酸甲酯

Reagent for stable and transient DNA/RNA transfections

同義詞:

N-(2,3-Dioleoyloxy-1-丙基)三甲胺甲基硫酸甲酯, 转染试剂

登入查看組織和合約定價


About This Item

經驗公式(希爾表示法):
C43H83NO8S
CAS號碼:
分子量::
774.19
MDL號碼:
分類程式碼代碼:
12352200
PubChem物質ID:

等級

for molecular biology

形狀

liquid (MES buffered saline)

用途

sufficient for 40 transfections (10 cm dishes)

濃度

1 mg/mL

顏色

clear to hazy colorless to faint yellow

儲存溫度

2-8°C

SMILES 字串

[H]C(C[N+](C)(C)C)(OC(CCCCCCC/C=C\CCCCCCCC)=O)COC(CCCCCCC/C=C\CCCCCCCC)=O.O=S([O-])(OC)=O

InChI

1S/C42H80NO4.CH4O4S/c1-6-8-10-12-14-16-18-20-22-24-26-28-30-32-34-36-41(44)46-39-40(38-43(3,4)5)47-42(45)37-35-33-31-29-27-25-23-21-19-17-15-13-11-9-7-2;1-5-6(2,3)4/h20-23,40H,6-19,24-39H2,1-5H3;1H3,(H,2,3,4)/q+1;/p-1/b22-20-,23-21-;

InChI 密鑰

RSMRWWHFJMENJH-LQDDAWAPSA-M

尋找類似的產品? 前往 產品比較指南

一般說明

DOTAP是一种由阳离子脂质体组成的化合物,用于向真核细胞中转染DNA、RNA及其他负电性分子。

應用

由阳离子脂质体组成的化合物,用于向真核细胞中转染DNA、RNA及其他负电性分子。DNA与DOTAP混合时会自动形成稳定的复合物,后者可直接加入细胞培养基中。这些复合物能够与细胞膜融合并向细胞质中释放DNA。细胞被高效转染的同时不会产生细胞毒性。每μg DNA需使用5-10 μg左右的DOTAP,每mL 细胞培养基需加入约10-20 μg DOTAP。最佳的工作浓度和转染试剂视被转染的细胞系,以及转染试材类型(RNA、DNA、蛋白)而定。
适用于向培养的哺乳动物细胞中瞬时或稳定转染DNA。每μg DNA需使用5-10 μg左右的DOTAP,每mL 细胞培养基需加入约10-20 μg DOTAP。根据使用指南对实验方案进行优化,能够获得最佳的转染效率。以下细胞系可通过DOTAP法成功转染:

A549
B6
BHK
BNL CL.2
C2
C6
心肌细胞
CHO
COS-1
COS-7
胚胎成纤维细胞(大鼠)
胚胎干细胞(小鼠)
成纤维细胞(人类原代细胞)
H225
肝细胞(大鼠原代细胞)
HepG2
HEK293
HeLa
海马神经元(小鼠与大鼠)
HMEC-1
Huh7
Jurkat
淋巴细胞(人类外周血)
M17
巨噬细胞(小鼠)
MCF-7
MDA-MB-435
NIH3T3
Sf9
平滑肌(大鼠主动脉)
肾脏(小鼠)
Vero
WI-38

特點和優勢

  • 转染效率比磷酸钙(沉淀法)与DEAE转染法高100倍
  • 细胞毒性比磷酸钙(沉淀)法与DEAE转染法更低
  • 同时兼容含血清和不含血清的转染实验
  • 同时适用于体内转染实验

原則

在不存在血清的条件下将DOTAP与DNA混合,即可形成稳定的复合物。该复合物性质稳定,可直接加入到细胞培养基中,并在培养基中与细胞膜融合,将DNA释放到细胞质中。注意:复合物的形成受到血清抑制,但一旦形成稳定的复合物,即便存在血清也无妨。

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 3

閃點(°F)

Not applicable

閃點(°C)

Not applicable

個人防護裝備

Eyeshields, Gloves


分析證明 (COA)

輸入產品批次/批號來搜索 分析證明 (COA)。在產品’s標籤上找到批次和批號,寫有 ‘Lot’或‘Batch’.。

已經擁有該產品?

您可以在文件庫中找到最近購買的產品相關文件。

存取文件庫

  1. 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.

  2. 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.

  3. 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.

  4. 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. 

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Lipid-Based Nanocarriers for RNA Delivery.
Xue HY
Current Pharmaceutical Design, 21(22), 3140-3147 (2015)
Venkat R Chirasani et al.
American journal of physiology. Cell physiology, 317(2), C358-C365 (2019-06-06)
Cryoelectron microscopy and mutational analyses have shown that type 1 ryanodine receptor (RyR1) amino acid residues RyR1-E3893, -E3967, and -T5001 are critical for Ca2+-mediated activation of skeletal muscle Ca2+ release channel. De novo missense mutation RyR1-Q3970K in the secondary binding
Thoria Donia et al.
Biochemical and biophysical research communications, 517(1), 146-154 (2019-07-29)
The phosphinositide PtdIns(3)P plays an important role in autophagy; however, the detailed mechanism of its activity remains unclear. Here, we used a Systematic Evolution of Ligands by EXponential enrichment (SELEX) screening approach to identify an RNA aptamer of 40 nucleotides
Lea I Mikkola et al.
PLoS genetics, 15(7), e1008197-e1008197 (2019-07-20)
Canine hip dysplasia is a common, non-congenital, complex and hereditary disorder. It can inflict severe pain via secondary osteoarthritis and lead to euthanasia. An analogous disorder exists in humans. The genetic background of hip dysplasia in both species has remained
Murali Muniraju et al.
Journal of virology, 93(16) (2019-05-31)
Kaposi sarcoma-associated herpesvirus (KSHV) is an emerging pathogen and is the causative infectious agent of Kaposi sarcoma and two malignancies of B cell origin. To date, there is no licensed KSHV vaccine. Development of an effective vaccine against KSHV continues

我們的科學家團隊在所有研究領域都有豐富的經驗,包括生命科學、材料科學、化學合成、色譜、分析等.

聯絡技術服務