Rat anti human il 6 antibodies

Supernatants were collected from HNECs and fibroblasts after 24 hours of exposure with different concentrations of deferiprone in the presence/absence of the pro-inflammatory agent Poly (I:C) (10 µg/ml)⁴⁰ or IL-1β (10 ng/ml Sigma, Saint Louis, USA)^{41 (link)} respectively. Interleukin-6 (IL-6) protein levels were estimated with an ELISA kit using rat anti-human IL-6 antibodies (BD Biosciences, New Jersey, USA), according to the manufacturer’s instructions. All measurements were performed in duplicate using a FLUOstar OPTIMA plate reader (BMG Labtech, Ortenberg, Germany). The tissue sample concentration was calculated from a standard curve and corrected for protein concentration.

Supernatant was collected from the basolateral compartment of treated HNEC-ALI cultures after 24 hours of exposure with inflammatory cytokines. Interleukin-6 (IL-6) protein levels were estimated with an ELISA kit using rat anti-human IL-6 antibodies (BD Biosciences, New Jersey, USA), according to the manufacturer’s instructions. All measurements were performed in duplicate using a FLUOstar OPTIMA plate reader (BMG Labtech, Ortenberg, Germany). The tissue sample concentration was calculated from a standard curve and corrected for protein concentration.

To determine if the chosen concentrations of antibiotics provoked an inherent inflammatory response likely to invoke a hypertrophic scar or adhesion, an IL-6 ELISA was undertaken on the cell supernatants. Supernatants were collected from fibroblasts and HNECs after 40 h of exposure to the antibiotics. Interleukin-6 (IL-6) protein levels were estimated with an ELISA kit using rat anti-human IL6 antibodies (BD Biosciences, New Jersey, USA), according to the manufacturer's instructions. All measurements were performed in duplicate using a FLUOstar OPTIMA plate reader (BMG Labtech, Ortenberg, Germany). The tissue sample concentration was calculated from a standard curve and corrected for protein concentration. These values were compared between antibiotic treatment groups, as well as scratched and unscratched control groups.