Biotin conjugated goat anti mouse igg

The following antibodies were used in this study: DCP1A (Santa Cruz, 56-Y), SYK (Santa Cruz, N19), G3BP1 (BD Biosciences, 611126), LC3-A/B (Cell Signaling Technology, D3U4C), p62 (Abcam, ab56416), E-cadherin for immunoblot (Santa Cruz, H10), DDX6 (Sigma Aldrich, P0067), GAPDH (Ambion, AM4300), SMAD2/3 (Cell Signaling Technology, 3102), phospho-SMAD2 (Cell Signaling Technology 3101), E-cadherin for IHC (BD biosciences, 610182), Ki67 (BD biosciences, 550609), AlexaFluor 594-conjugated goat anti-rabbit IgG (Invitrogen), AlexaFluor 594-conjugated goat anti-mouse IgG (Invitrogen), and AlexaFluor 488-conjugated goat anti-mouse IgG (Invitrogen), biotin-conjugated goat anti-mouse IgG (Jackson). For immunohistochemistry biotinylated secondary antibodies were detected using the ABC elite kit in combination with 3-3′-diaminobenzidine (Vector).

Influenza-specific antibodies were quantified by ELISA, whereby MaxiSorp ELISA plates (Nunc) were coated overnight at 4°C with UV-inactivated PR/8 (4000 PFU/ml). After blocking of unspecific binding (PBS 1% BSA), 50 μl of 1: 12500 diluted serum samples were added, followed by detection antibody (biotin-conjugated goat anti-mouse IgG, Jackson ImmunoResearch) and horseradish peroxidase-conjugated streptavidin (BD Pharmingen). Signal emitted by TMB liquid substrate (Sigma) was detected at 450 nm (620 nm reference) using a Sunrise plate reader (Tecan).