Clone mq2 13a5

To quantify human IL-6 and IFN-β production, specific capture ELISAs were performed. A rat anti-human IL-6 capture antibody (BD Pharmingen, cat# 554543; Clone Mq2-13A5, 2 μg/ml) and a biotinylated rat anti-human IL-6 detection antibody (BD Pharmingen, cat# 554546; Clone MQ2-39C3, 2 μg/ml) were used in IL-6 ELISAs. While, a polyclonal rabbit anti-human IFN-β capture antibody (Abcam, cat# ab186669, 0.25 μg/ml) and a biotinylated polyclonal rabbit anti-human IFN-β detection antibody (Abcam, cat# ab84258, 0.25 μg/ml) were used in IFN-β ELISAs. Bound antibody was detected using streptavidin-HRP (BD Biosciences) followed by the addition of tetramethylbenzidine substrate. H₂SO₄ was used to stop the reaction and absorbance was measured at 450 nm. A standard curve was generated using dilution of recombinant cytokines for IL-6 (BD Pharmingen) and IFN-β (Abcam). The cytokine concentration in cell supernatants was determined by extrapolation of absorbance to the standard curve.

Specific capture ELISA were performed to quantify human IL-6 and IFN-β production. A rat anti-human IL-6 capture antibody (BD Pharmingen, cat# 554543, Clone Mq2-13A5) and a biotinylated rat anti-human IL-6 detection antibody (BD Pharmingen, cat# 554546, Clone MQ2-39C3) were used for IL-6 capture ELISAs. A polyclonal rabbit anti-human IFN-β capture antibody (Abcam, cat# ab186669) and a biotinylated polyclonal rabbit anti-human IFN-β detection antibody (Abcam, cat# ab84258) were used in IFN-β capture ELISAs. Streptavidin–horseradish peroxidase (HRP) (BD Biosciences) was added prior to the addition of tetramethylbenzidine substrate to detect bound antibody. The reaction was stopped using H₂SO₄ and the absorbance was measured at 450 nm. Recombinant cytokines for IL-6 (BD Pharmingen) or IFN-β (Abcam) were diluted to generate standard curves and the cytokine concentration in cell supernatants was determined by extrapolation of absorbance to the standard curve.