Goat anti human igg hrp antibody

Total serum IgA1 and IgA2 levels were measured by in-house ELISA as previously described.11 (link) Briefly, ThermoFisher Nunc MaxiSorp flat-bottom plates were precoated with 2 µg/mL of goat F(ab)2 anti-human IgG (H+L) (Jackson ImmunoResearch #109-006-003) or goat F(ab)2 anti-human IgA (SouthernBiotech, #2052-01) overnight at 4°C. On the next day, plates were washed with 0.05% PBS/Tween and blocked with 1% PBS/bovine serum albumin (BSA). Sera (diluted 1:1000 000 for IgG and 1:40 000 for IgA measurement) and IgG commercial standard (Human IgG I2511 Sigma-Aldrich) or isolated IgA in known concentrations were incubated on the plates for 1 hour. After the plates were washed, they were incubated with goat anti-human IgG HRP-antibody (SouthernBiotech #2040-05) for 1 hour. After detection with TMB substrate, the absorption was measured at 450 nm with reference wavelength at 650 nm.