Human il 6 quantikine hs elisa high sensitivity kit

Interleukin-6 (IL-6), C-reactive protein (CRP), serum amyloid A (SAA), fibrinogen, factor VIII, von Willebrand factor, and Clara cell protein (CC16) were analyzed in venous plasma or serum collected before, 3.5 and 22 h post-exposure. CC16 and IL-6 were analyzed using the Human Uteroglobin DuoSet® ELISA Developmental system and Human IL-6 Quantikine® HS Elisa High Sensitivity kit both from R&D Systems® (Minneapolis, MN) respectively according to the protocol by the manufacturer. The LODs were 7.81 pg/ml and 0.016 pg/ml for CC16 and IL-6, respectively.

Sputum induction and processing were performed at 6 h from the start of exposure in control and high ACR exposure as previously described (Strandberg et al., 2008 (link)). Briefly, the sputum weight was determined and an equal volume of dithiothreitol 0.1% (Sputolysin® reagent, Calbiochem, San Diego, CA) was added to the whole sputum sample and rocked for 15–20 min in a 37 °C in a shaking water bath. The sample was centrifuged (10 min at 280 g). The cell pellet was re-suspended in 2 ml phosphate-buffered saline and put on ice. Total cell count and viability tests with Türk (Histolab Products AB, Gothenburg, Sweden) and Trypan blue (Sigma Aldrich, Steinheim, Germany) were performed. Cytocentrifuge-prepared slides were stained with May-Grünwald Giemsa stain (Sigma Aldrich, Steinheim, Germany) and 300 cells (squamous cells excluded) were assessed for differential cell counts. The sputum samples were included in the analysis if they contained less than 30% squamous cells. The supernatant was dispensed into aliquots and kept in −70 °C until analysis. IL-6 and interleukin-8 (IL-8) were analyzed in supernatant of the sputum. They were analyzed by using Human IL-6 Quantikine® HS Elisa High Sensitivity kit and Human CXCL/IL-8 duo set from R&D Systems® (Minneapolis, MN). The LOD were 0.016 pg/ml and 31.2 pg/ml for IL-6 and IL-8, respectively.