Any advise on the ideal dilution factor to use for culture supernatants sample preparation for multiplex assay? I'm planning to run a multiplex assay experiment for the detection of cytokine/chemokine stimulation, but struggling with the right dilution factor of my culture supernatants samples harvested from an M. tuberculosis infected raw macrophages cell line.The Bio- Plex Pro manual only gives a dllution factor for plasma sample type but not culture suparnatants instead this is mostly user defined. Any suggestions or papers recommendation?
I have detected cytokines in supernatant samples before; we did not need to dilute any of the samples; dilution was important for antibodies, which is usually given as a range (quite wide) on the datasheet from the supplier. I can loosely suggest that you adopt the dilution that is specified for plasma samples (if there is room for trial/error). From my experience, a little tweaking of the dilutions is necessary to optimise detection.
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