Biotinylated anti mouse ige ab

Total IgE Ab levels were determined by a BD OptEIA Set Mouse IgE, (BD PharMingen) according to instructions from the manufacturer. To prevent interference of IgG in the assay, serial dilutions of immune plasma were previously depleted of IgG by overnight incubation in Reacti-Bind Protein G coated plates (Pierce) (38 (link)). Antigen-specific IgE was detected with a biotinylated anti-mouse IgE Ab (BD Biosciences).

Total IgE Ab levels were determined by a BD OptEIA™ Set Mouse IgE (BD PharMingen) according to instructions from the manufacturer. To prevent interference of IgG in the assay, serial dilutions of immune plasma were first depleted of IgG by overnight incubation in Reacti-Bind™ Protein G Coated Plates (Pierce, Rockford, IL, USA). In order to detect antigen-specific IgE, the microtiter plates were coated with OVA (1 mg/ml). Serial dilutions of IgG-depleted plasma were then added and IgE were detected with a biotinylated anti-mouse IgE Ab (BD Biosciences). The IgE titers were determined as the last dilution of samples that achieved an absorbance of >0.1 above that of control samples.