Thursday, July 18, 2013

Applications Thursday: Using the FLUOstar Optima for the Detection of Lonza's Kinetic Endotoxin Kit

Endotoxins or lipopolysaccharides (LPS) often contaminate plasmid DNA and protein preparations that are isolated from gram-negative bacteria. This contamination can have serious consequences as even small amounts of LPS can lead to an inflammatory response resulting in sepsis in a condition called endotoxemia. LPS must, therefore, be detected and eliminated if the DNA or protein are to be used in in vivo applications like gene therapy.

The BMG LABTECH application note entitled: 'Lonza’s Kinetic Kit for Endotoxin Detection Using the FLUOstar OPTIMA and MARS Data Analysis' describes an approach that allows users to confidently detect LPS contamination. Lonza's kinetic kit contains a proenzyme that is activated in the presence of endotoxin which cleaves a colorless peptide to release p-nitroaniline which is detectable at 405 nm.
Signal curves for several endotoxin samples taken directly
from MARS data analysis software.

Kinetic signal curves for standards and unknown could then be produced and analyzed in the MARS data analysis software that is provided with all BMG LABTECH microplate readers. The result of the analysis is a standard curve that can be used to reliably determine the concentration of LPS in the unknown samples.

For the details of this and other applications suitable for use on BMG LABTECH microplate readers please visit the Applications Center at our website:

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