Ju Yan-fang, Yang Jin-ju, Liu Rong, Gao Yuan, Liu Xiao-lan, Gao Jian-en, Sun Qi-hong
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 May;22(3):377-9.
To prepare monoclonal antibodies (mAbs) against Dicarbonyl/L-xylulose reductase (DCXR).
Normal human liver tissues were homogenized, and mitochondria were isolated by differential centrifugation. The total mitochondrial proteins were used to immunize BALB/c mice to prepare mAbs by routine hybridoma technique. The mAbs were detected by ELISA, Western blot and immunohistochemistry. The antibody specificities were identified by mass spectrometry (MS) following immunoprecipitation (IP) and confirmed by Uni-ZAP expression library screening.
One clone of hybridoma AFF091 secreting specific mAb against DCXR was obtained. The Ig subclass of the mAb was IgG1 and it can be used in ELISA, Western blot, immunohistochemistry, and immunoprecipitation.
A hybridoma cell line stably secreting specific mAb against DCXR was established. The specific mAb against DCXR would be very useful for investigation of DCXR functions and distribution.
制备抗二羰基/L-木酮糖还原酶（DCXR）的单克隆抗体（mAb）。
将正常人肝组织匀浆，通过差速离心分离线粒体。用总线粒体蛋白免疫BALB/c小鼠，采用常规杂交瘤技术制备单克隆抗体。通过酶联免疫吸附测定（ELISA）、蛋白质印迹法（Western blot）和免疫组织化学法检测单克隆抗体。在免疫沉淀（IP）后通过质谱（MS）鉴定抗体特异性，并通过Uni-ZAP表达文库筛选进行确认。
获得了一株分泌抗DCXR特异性单克隆抗体的杂交瘤AFF091。该单克隆抗体的Ig亚类为IgG1，可用于ELISA、Western blot、免疫组织化学和免疫沉淀。
建立了稳定分泌抗DCXR特异性单克隆抗体的杂交瘤细胞系。抗DCXR的特异性单克隆抗体对于研究DCXR的功能和分布将非常有用。
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