Interleukin-1 is a pro-inflammatory cytokine that can be expressed in two separate isoforms: IL-1α and IL-1β.1 IL-1β is expressed by dendritic cells and acts through the TNFα signaling pathway to induce expression of several inflammatory cytokines2, 3; it has therefore been identified as both a biomarker for inflammation diagnosis and a target for inflammation therapy.
Current ELISAs lack the required sensitivity to quantify IL-1β in normal healthy donor serum for proper comparison with diseased individuals. Sword has changed that by developing a reagent based on the principle of Raman Resonance that can easily be inserted into a standard IL-1β ELISA with greatly enhanced sensitivity.
This Sword Assay has been optimized for use with the R&D Systems DuoSet ELISA for Human IL-1β (Catalog No. DY201). This protocol allows the user to use 50 µL sample sizes instead of 100 µL as specified in the DuoSet literature.
Table 1. Precision of IL-1β Quantification in Human Serum with Sword Assay
Table 1. IL-1β levels were quantified in human serum from healthy donors using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β. Donor samples were tested in duplicate in three separate runs.
Table 2. Precision of IL-1β Quantification in Human Plasma EDTA with Sword Assay
Table 2. IL-1β levels were quantified in human plasma EDTA from healthy donors using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β. Donor samples were tested in duplicate in three separate runs.
Recovery
Table 3. Spike Recovery with Sword Assay for Human IL-1β in Serum
Table 3. Human IL-1β Reference Standard was spiked into pooled human serum from healthy donors. Human IL-1β levels were quantified using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β.
Table 4. Spike Recovery with Sword Assay for Human IL-1β in Plasma EDTA
Table 4. Human IL-1β Reference Standard was spiked into pooled human plasma EDTA from healthy donors. Human IL-1β levels were quantified using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β.
Quantification
Table 5. Quantification of IL-1β in Healthy Human Serum
Table 5. Human IL-1β was quantified in human serum from twelve healthy donors using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β.
Table 6. Quantification of IL-1β in Healthy Human Plasma
Table 6. Human IL-1β was quantified in human plasma from twelve healthy donors using the R&D Systems Human IL-1β DuoSet ELISA (DY201) with Sword Assay for Human IL-1β.
Citations
Weber A, et al: Interleukin-1beta (IL-1beta) processing pathway. Sci Signal 2010, 3:cm2.
Luft T, et al: IL-1 beta enhances CD40 ligand-mediated cytokine secretion by human dendritic cells (DC): a mechanism for T cell-independent DC activation. Journal of immunology 2002, 168:713-22.
Zhang et al: Interleukin-1beta induces macrophage inflammatory protein-1beta expression in human hepatocytes. Cell Immunol 2003, 226:45-53.
Ready to Optimize Your Assay?
Put Sword to the test. Give us 6 days and we’ll show you just how much we can boost your specific assay’s sensitivity, precision, and accuracy. We’ll even assess your first target assay free of charge.