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Endotoxins, also known as lipopolysaccharides (LPS), are large molecules (molecular weight 200-1000 kDa) present on the outer membrane of Gram negative bacteria. The outer membrane provides a barrier to protect cells from damage from large molecules and hydrophobic compounds, but hydrophilic small molecules crucial for cell survival are permeable. Endotoxins may also play a major role in host cell interactions, such as colonization and resistance to phagocytosis. Endotoxins are composed of polysaccharide regions and a lipid region (lipid A), which are responsible for inducing toxicity. Endotoxins are highly toxic to humans and may lead to various life-threatening diseases. Endotoxins obtained from the environment can cause respiratory symptoms, lung inflammation, and asthma.
Endothelin is released in small amounts from cells during cell growth, and in large amounts during cell death and lysis, thus also during plasmid purification. Like intact cells, free LPS molecules induce inflammatory responses in the mammalian immune system. Therefore, it is necessary to quantitatively remove them from plasmid formulations to ensure high transfection rates and high viability of transfected cells. Like DNA, endotoxins are negatively charged and co-purified with plasmid DNA in silica-based and anion-exchange-based systems.
Plasmids separated using traditional purification procedures typically contain high levels of endotoxin (also known as lipopolysaccharide or LPS), which can significantly interfere with downstream transfection experiments. Our Endo-Free Plasmid DNA Purification Kits integrate effective endotoxin removal steps into the plasmid purification program to produce high-quality transfection grade (<1 EU/µg) plasmids for efficient transfection. The working procedure for preparing endotoxin free plasmids is the same as the standard protocol, and does not require time-consuming cultivation steps to remove endotoxins.
| Cat.No. | Product Name | Price |
|---|---|---|
| NKP-T018 | Plasmid Endo-Removal Ultrapure Kit | Inquiry |
| NKP-T019 | Plasmid Endo-Free Ultrapure Kit | Inquiry |
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