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I have purchased Listeriolysin O (LLO) protein (information as attached) and was found that it is not active for some reason! I was trying to measure the conduction of the protein by using impedance spectroscopy and tethered bilayer lipid membranes (with the use of up to 50 mol% cholesterol and other neutral lipids) but actually I have tried different concentrations of the protein ranging from 400nM to 52uM and still the protein did not show any kind of activity which is very unusual because the system was tested with the use of different toxins and proteins including alpha hemolysin and CLIC1.
Any help will be highly appreciated to get the protein working.
Please refer to below for further details:
Listeriolysin O - Cat No ab83345
Asked on Jul 04 2012
Thank you for contacting us. I am sorry to hear that this protein is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.
Having reviewed this case, our collaborators from the lab where the protein is producedoffered some suggestions to help optimize the results from ab83345:
"The procedures require that the hemolytic activity of each new batch of listeriolysin O PEST-minus is determined by the assay described in Giammarini C. et al. "Purification and characterization of a recombinant listeriolysin O expressed in Escherichia coli and possible diagnostic applications", .J Biotechnol. 2004 Apr 8;109(1-2):13-20.
In particular, listeriolysin O PEST minus lotGR61074 has provided the result of 7x10(4) HU/mg, that corresponds to 1HU = 14.2ng.
Unfortunately we have no information on the performance of our product in the specific application described by the customer".
Should the suggestions not improve the results, please do let me know.
In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.
I hope this information is helpful, and I thank you for your cooperation.
Answered on Jul 04 2012