Immunohistochemical (IHC) analysis (Fig. 6c and Supplementary Fig. 4) was performed as previously described2 (link),17 (link),26 (link),31 (link),35 using an Autostainer Link 48 (Dako). The deparaffinized sections were exposed to EnVision FLEX target retrieval solution high pH (Agilent, Cat# K8004) for 20 min at 97°C for activation, and a mouse anti-SARS-CoV-2 N monoclonal antibody (clone 1035111, R&D Systems, Cat# MAB10474-SP, 1:400) was used as a primary antibody. The sections were sensitized using EnVision FLEX for 15 min and visualized by peroxidase-based enzymatic reaction with 3,3’-diaminobenzidine tetrahydrochloride (Dako, Cat# DM827) as substrate for 5 min. The N protein positivity was evaluated by certificated pathologists as previously described2 (link),17 (link),26 (link),31 (link),35 . Images were incorporated as virtual slides by NDP.scan software v3.2.4 (Hamamatsu Photonics). The N-protein positivity was measured as the area using Fiji software v2.2.0 (ImageJ).
Ito J., Suzuki R., Uriu K., Itakura Y., Zahradnik J., Kimura K.T., Deguchi S., Wang L., Lytras S., Tamura T., Kida I., Nasser H., Shofa M., Begum M.M., Tsuda M., Oda Y., Suzuki T., Sasaki J., Sasaki-Tabata K., Fujita S., Yoshimatsu K., Ito H., Nao N., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Yamamoto Y., Nagamoto T., Kuramochi J., Schreiber G., Saito A., Matsuno K., Takayama K., Hashiguchi T., Tanaka S., Fukuhara T., Ikeda T, & Sato K. (2023). Convergent evolution of SARS-CoV-2 Omicron subvariants leading to the emergence of BQ.1.1 variant. Nature Communications, 14, 2671.
Anti antibody Antibody Clone Enzymatic Mouse N proteinPathologistsPeroxidase Sars cov 2 Scan
Corresponding Organization : National Institute of Infectious Diseases
Other organizations :
Hokkaido University, Weizmann Institute of Science, Kyoto University, MRC University of Glasgow Centre for Virus Research, Medical Research Council, Kumamoto University, University of Miyazaki, Kyushu University, Tokyo Metropolitan Institute of Public Health, Saiseikai Utsunomiya hospital, University Children's Hospital Tübingen, Tokai University, Hiroshima University, Japan Agency for Medical Research and Development
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