Mini elisa plate reader

Cytokine and chemokines involved in neutrophil mobilization and activation were measured using a custom multiplex bead array kit (LEGENDPlex, Biolegend, San Diego, CA, USA). Twelve cytokines were measured in duplicate, including CXCL1, CXCL2, G-CSF, IL-1b, IL-6, IL-4, TGFb, TNFa, GM-CSF, INFg, IL-17A, and IL-10. The assay was performed according to manufacturer’s instructions, collected on an SH800 (Sony), and analyzed using the LEGENDPlex Cloud-Based Data Analysis Software version 2023-02-15 (Biolegend, San Diego, CA, USA and Qognit, San Carlos, CA, USA). Neutrophil elastase was measured via ELISA using a pre-validated kit following manufacturer’s instructions (Abcam, Cambridge, UK) and read using a Mini ELISA Plate Reader (Biolegend, San Diego, CA, USA).

Cytokine and chemokines involved in neutrophil mobilization and activation were measured using a custom multiplex bead array kit (LEGENDPlex, Biolegend). Twelve cytokines were measured including, CXCL1, CXCL2, G-CSF, IL-1b, IL-6, IL-4, TGFb, TNFa, GM-CSF, INFg, IL-17A, and IL-10. The assay was performed according to manufacturer’s instructions in duplicate, collected on an SH800 (Sony), and analyzed using the LEGENDPlex Cloud-Based Data Analysis Software (Biolegend and Qognit). Neutrophil elastase was measured via ELISA using a pre-validated kit following manufacturer’s instructions (Abcam) and read using a Mini ELISA Plate Reader (Biolegend).

IFN-γ was measured using the IFN-γ ELISA MAX Deluxe Set (BioLegend, catalogue #430116) and performed according to the manufacturer’s instructions. Immediately following the addition of stop solution (2 N H₂SO₄), microplates were read at 450 nm using the BioLegend Mini ELISA Plate Reader. IFN-γ was quantified by extrapolating from the standard curve using GraphPad Prism. Values below the lower limit of detection of the assay were recorded as 7.8 pg/ml; values above the upper limit of detection of the assay were recorded as 1000 pg/ml.