Rabbit anti mouse iga antibody

To determine the specific IgA, the interior of the small intestine from immunized mice was washed twice with a total of 1 ml of cold PBS. After centrifugation at 800 g for 10 min, the supernatants of the intestinal washes were harvested. The specific anti-TsNd IgA was measured by standard ELISA using an rTsNd-coated plate. Total IgA was quantified by a sandwich ELISA using rabbit anti-mouse IgA antibody (Abcam, UK) as the capture antibody and HRP-conjugated goat anti-mouse IgA antibodies as the detection antibody. To compensate for variations in the efficiency of the recovery of secretory IgA antibodies among animals, the antigen-specific IgA in each sample was normalized to the total IgA present in the lavage.

Mycobacterial loads in lungs and spleen of Mtb-infected mice were determined at different time points post-infection as previously described (29 (link)). Lungs of Mtb-infected mice were fixed with 4% phosphate-buffered formalin, and 3 μm-thick sections were stained with either H&E or rabbit anti-mouse antibody specific for iNOS (Abcam) or rabbit anti-mouse IgA antibody (Abcam). Detection was performed using HRP-labelled anti-rabbit antibody (Dako) followed by 3, 3’-diaminobenzidine substrate (Dako). The lung images and lesion areas, iNOS and IgA positive areas were acquired in Nikon 90i Eclipse widefield microscope and quantified using NIS elements.