• Eri Sata, Leo Takada, Ryo Kaetsu, Mai Fukasawa, Mariko Ohtsu, Mitsuru Motoyoshi, and Masatake Asano.
• Department of Orthodontics, Nihon University School of Dentistry.
• J Oral Sci. 2020 Jun 23; 62 (3): 340-343.

AbstractInterleukin-1α (IL-1α) is produced inside cells in its precursor form (pIL-1α). Enzymatic cleavage yields mature (mIL-1α) and the propiece of IL-1α (ppIL-1α), which are thought to be localized in the nucleus, because of the presence of nuclear localizing signals. Studies of ppIL-1α function have been hampered by the lack of a ppIL-1α-specific antibody (Ab). In the present study, the authors generated anti-ppIL-1α Ab by using recombinant histidine-tagged ppIL-1α (His-ppIL-1α) as an immunogen. Rabbits were immunized with His-ppIL-1α, and affinity-purified Ab was obtained. Ab reactivity and specificity were examined by Western blotting. The antibody successfully recognized transfectant-derived green fluorescence protein (GFP)-tagged ppIL-1α but not GFP. A sandwich enzyme-linked immunosorbent assay (ELISA) system established by biotinylating the anti-ppIL-1α Ab successfully detected GFP-ppIL-1α. The Ab and ELISA system allows functional analysis of ppIL-1α and improves understanding of ppIL-1α.

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