Hello everyone,
I was wondering if anyone has used Pierce™ Chromogenic Endotoxin Quant Kit to detect endotoxin presence of a powder sample diluted in DMSO.
Firstly, we are not sure whether our samples contain endotoxin, so we do not where to start since the kit procedure contains high and low standards for quantification, dilutions of which depends on the amount of endotoxin present in the samples.
Secondly, the kit says if samples are colored (ours are dark yellow), this may interfere with the results so we should dilute our samples. However, again since we are not sure whether our samples contain endotoxin or how much dilution would be feasible to use for the detection range of the kit, we do not know where to start.
Does anyone have any experience using this kit in an unknown sample? Any suggestions you would give is highly appreciated.
Thanks
Asli
If you already have the kit, then I would suggest trying it out. You would need three assay groups: first is your reconstituted sample diluted serially, the second group is your sample spiked with a known amount of LPS (endotoxin; use the stock solution that you would use to create your standards), and the third is DMSO neat (to make sure your solvent doesn't have LPS). Run that and see what you see.
If the powder is the bottleneck due to cost/production then I would suggest using a turbidimetric assay or a fluorescence based assay. Make sure that DMSO is not an inhibiting reagent for whatever test you get.
Hello Asli Korkmaz ,
Sorry, my message will not shed light on your question, but I hope that you have finally been able to analyse your samples.
Actually, I am ready to order a Pierce™ Chromogenic Endotoxin Quant Kit. I am not sure yet which version of the kit to order, the 60 or the 240 reactions? Knowing the shelf life of the kit will determine my choice.
Do you know how long it can be used once opened?