Mouse ifnγ elisa max kit

Cytokines from serum, liver homogenate, and cell culture supernatants were measured using the following ELISA or Luminex kits according to the manufacturer’s instructions: Mouse-IFNγ ELISA MAX Kit (BioLegend), MILLIPLEX Mouse Th17 Magnetic Bead Panel (Millipore Sigma), Cytokine & Chemokine 36-Plex Mouse Procarta Plex Panel 1A (Thermo Fisher Scientific-Affymetrix), and IL-18 Mouse ELISA Kit (Invitrogen).

Interferon gamma was detected using the “Mouse IFN-γ ELISA MAX” kit from Biolegend (#430805) in accordance with manufacturer’s instructions. Detection of anti-EBOV GP antibodies was performed by coating optical plates overnightwith soluble EBOV GP (50 ml/well at 10 mg/ml). Wells were washed 2x with PBST (0.015% Tween 20), blocked for 1 hour (PBS 2%BSA), washed 3x with PBST, and incubated overnight in 4°C with either a standard curve composed of fractionated mouse immunoglobulin (Immunore-agents #Mu-003-B) or serum samples. Following incubation wells were washed 4x with PBST and incubated with 50 mL rabbit anti-mouse IgG heavy and light chain-HRP (10 mg/ml Pierce #31457) for 1 hr at RT. Wells were then washed 5x with PBST, incubated with 50 mL of HRP substrate (BD OptEIA #555214). The reaction was stopped with 50 mL 2M H₂SO₄ and absorbance at 450nm was measured on Synergy H1 hybrid reader. Total concentration of anti-EBOV GP antibodies was quantified by comparison to the standard curve.