Bio-Rad Laboratories has announced the availability of a technical note (5762) that describes a human cytokine study undertaken by researchers using the company's Bio-Plex suspension array system.
For the study, the researchers at the Institute of Cell Biology in Zurich, Switzerland, developed an RNAi-based method for analysing caspase-1-dependent release of various cytokines.
The Bio-Plex immunoassay panel used measured IL-1B in addition to 26 other cytokines, 21 of which possessed a signal peptide.
This allowed them to determine whether caspase-1 specifically targets unconventional protein secretion or secretion in general.
The results showed that it is critical to measure cytokine concentrations in both supernatant and cell lysate when measuring secretion because it allows the calculations of percentages of secretion, thereby accounting for changes in protein expression.
The researchers were able to verify that in keratinocytes, caspase-1 only affects unconventional protein secretion and not protein expression in general.
The researchers concluded that their approach can be applied to virtually all cell types that can be cultured, as well as to tissue cultures.
SiRNA, specific inhibitor, and knockdown cells can then be used to investigate the impact of genes or cellular processes on protein secretion.
The researchers also determined that multiplex cytokine suspension array technology is a powerful tool for identifying cytokines that are secreted from cells under certain conditions.
The key advantage of this method is the associated quantitative measurement of several cytokines in the same experiment.
