UltraClean® Endotoxin Removal Kit

Overview

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Description

The UltraClean^® Endotoxin Removal Kit is designed to remove endotoxins from previously isolated plasmid DNA in low salt solutions. Solutions containing high salt concentrations should be diluted to 100 mM or less before using this kit. Plasmid DNA is mixed with the UltraClean^® Endotoxin Removal Reagent and briefly incubated on ice. The mixture is then heated followed by centrifugation. Plasmid DNA remains in the aqueous phase while endotoxins are removed in the organic phase of the extraction. Multiple rounds of extraction are recommended for plasmid DNA to ensure complete removal of contaminating endotoxins.

If the extracted DNA is too dilute, it can be concentrated using the Endotoxin-Free Precipitation Reagents included in this kit (Ethanol, Water and 5 M NaCl). After treatment with this kit, the plasmid DNA will have endotoxin levels that are less than 0.1 EU/µg. The treated plasmid DNA is then suitable for transfection, injection and other sensitive applications.

Endotoxins, also known as lipopolysaccharides, are cell membrane 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. Endotoxins represent a non-controllable variable in transfection experiment setup, making results difficult to compare and interpret.

MSDS

12615

Protocol

12615

Specifications

Storage Conditions

Publications

• Laroche-Pierre, S., Jodoin, J., and LeBlanc, A.C. Helix 2 is Necessary and Sufficient for Prion Protein’s Anti-Bax Function. Journal of Neurochemistry. 108(4): 1019-1031 (2009).
• Lin, D.T.S., Jodin, J., Baril, M., Goodyer, C.G., LeBlanc, A.C. Cytosolic Prion Protein is the Predominant Anti-Bax Prion Protein Form in the Anti-Bax Function. Biochimica et Biophysica Acta (BBA). 1783(10): 2001-2012 (2008).
   

Kit Components