Goat anti mouse il 33 igg

Tumors were surgically removed and fixed in 10% phosphate-buffered formalin solution. The tissues were embedded in paraffin and cut into 2-μm sections. After heat-induced antigen retrieval in REAL Target Retrieval Solution (DAKO) and subsequent blocking of nonspecific sites with 0.1% normal goat serum/1% BSA, the tissues were immunostained with rabbit polyclonal anti-myeloperoxidase (MPO) antibody (Bioss Inc., Boston, MA, USA) diluted 1:200 using a previously described method [22 (link)]. The tissues were also immunostained with rabbit polyclonal anti-MPO antibody or rabbit polyclonal anti-CD20 antibody (Thermo Fisher Scientific) diluted 1:200, followed by Alexa Fluor 594-conjugated goat anti-rat IgG or phycoerythrin (PE)-conjugated rat monoclonal anti-mouse F4/80 (Abcam) diluted 1:100. In some experiments, after the tissues were treated with M.O.M. (Mouse on Mouse) Immunodetection Kit-Fluorescein (Vector Laboratories, Inc., Burlingame, CA, USA), the tissues were double immunostained with 10 μg/ml goat anti-mouse IL-33 IgG (R&D Systems, Minneapolis, MN, USA) and mouse monoclonal anti-α-SMA antibody diluted 1:100 (DAKO), followed by Alexa Fluor 594-conjugated chicken anti-goat IgG and Alexa Fluor 488-conjugated goat anti-mouse IgG. The sections were treated with the Vector TrueVIEW Autofluorescence Quenching Kit (Vector Laboratories) and counterstained with DAPI.