Interleukin-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.

白细胞介素-1α（IL-1α）以其前体形式（pIL-1α）在细胞内部产生。酶促裂解产生成熟的（mIL-1α）和IL-1α的前体（ppIL-1α），由于存在核定位信号，它们被认为位于细胞核中。缺少ppIL-1α特异性抗体（Ab）阻碍了ppIL-1α功能的研究。在本研究中，作者通过使用重组组氨酸标签的ppIL-1α（His-ppIL-1α）作为免疫原产生了抗ppIL-1αAb。用His-ppIL-1α免疫兔子，获得亲和纯化的Ab。通过蛋白质印迹检查抗体的反应性和特异性。该抗体成功识别了转染子来源的绿色荧光蛋白（GFP）标签的ppIL-1α，但未识别GFP。通过对抗ppIL-1αAb进行生物素化而建立的三明治酶联免疫吸附测定（ELISA）系统成功检测到GFP-ppIL-1α。Ab和ELISA系统可对ppIL-1α进行功能分析，并增进对ppIL-1α的了解。