Good-bye Endotoxin, Hello Transfection

Norgen’s Endotoxin Removal Kit (Mini) is designed for the rapid removal of endotoxins from up to 40 µg of previously purified DNA. Endotoxins are cellmembrane components of Gram-negative bacteria such as E. coli. Endotoxins are released during the lysis step of plasmid purification and significantly reduce transfection efficiencies in endotoxin sensitive cell lines. Therefore, the removal of endotoxins from plasmid preparations is often necessary prior to the use of the DNA in downstream applications. With Norgen’s Endotoxin Removal Kit (Mini) endotoxin levels are efficiently reduced to 0.1 EU/ µg DNA or less. Company Website

Endotoxins, or lipopolysaccharides, co-purify with DNA on ion exchange and size exlusion columns. Because endotoxins are extremely toxic to primary cells, this becomes an obstacle to in vitro and in vivo transfection. Sigma’s Endotoxin removal reagent relies on the micellar properties of Triton X-114 to remove endotoxins from solution. When warmed above 20 °C, micelles aggregate into a separate phase that sequesters amphiphilic solutes, including lipopolysaccharides. The hydrophilic DNA remains in the aqueous phase. Company Website

Pierce's Detoxi-Gel provides a quick method for removal of endotoxins from samples. This versatile gel reduces endotoxin levels in protein solutions, cell culture media, solutions containing pharmacological components, and aqueous buffers. This technology utilizes an immobilized ligand to bind and remove pyrogens, and the endotoxin removal efficiency is greater than 99%. The Detoxi-Gel can be regenerated by stripping off the endotoxins with a 1% deoxycholate solution in pyrogen-poor water, and it suffers no loss in binding capacity after 10 regenerations. Company Website