Polyclonal goat igg anti human kappa

Female Balb/c and male NSG mice were injected intravenously (i.v.) with 5 mg/kg antibody solution in PBS and blood was sampled from the submandibular vein at indicated time points. Samples were left to coagulate overnight in Eppendorf tubes at 4°C. After centrifuging twice at 3000 × g, serum was collected and stored at −20°C. MaxiSorp 96-well plates were coated with 0.5 µg/mL polyclonal goat-IgG anti-human kappa (1:2000, SouthernBiotech) in PBS overnight at 4°C. Next, plates were washed thrice with wash buffer (0.05% Tween20 in PBS) and blocked for 1 hour with 1% BSA in wash buffer. Serum samples and standards were diluted in PBS and incubated for 1.5 hours at RT. After washing thrice, polyclonal goat anti-human IgA/IgG-HRP (1:2000, SouthernBiotech) was added for 1 hour. After washing thrice again, plates were developed for 10 min with ABTS (Roche) and measured on a spectrophotometer (Bio-Rad) at 415 nm.

MaxiSorp 96 well ELISA plates (NUNC, Thermo Fisher) were coated overnight with 0.5 µg/mL polyclonal goat IgG anti-human kappa (Southern Biotech) diluted in PBS. Next, plates were washed three times with 0.05% TWEEN 20 in PBS (PBST) and blocked for 1 hour by incubating with 1% BSA (Roche) in PBST. Serum samples were diluted 1:2000 in 1% BSA in PBST and incubated for 1.5 hours at room temperature. Next, plates were washed three times with PBST. HRP-labeled-anti-human IgA or HRP-labeled-anti-human-IgG (SouthernBiotech) were used to bind human IgA or human IgG, respectively. Plates were developed for 10 min with ABTS (Roche) and read out on a spectrophotometer (Bio-Rad) at 415 nm.