Quantification of IgE and IgG Levels

The total IgE level was determined in the mouse serum using MILLIPLEX^® MAP Mouse IgE Single Plex Magnetic Bead Kit (Merck Millipore, Banglore, India) as per the manufacturer’s protocol and the samples were analyzed using the Luminex Magpix (Merck Millipore, Banglore, India) [22 (link)]. The total IgG level in the mouse serum was measured using the Amplex ELISA development kit (San Francisco, CA, USA) for mouse IgG as per the manufacturer’s protocol. All the samples were analyzed at 405 nm using ELISA plate reader. The results obtained were means of duplicate determination with variation less than 10%. Further, the IgE and IgG concentrations were determined by comparing the mean OD values of the tested sera with the mean OD values of the standard. IgE and IgG titers were calculated by multiplying the dilution factor of the test sera and expressed in ng/mL.