[Preparation and characterization of monoclonal antibody against DCXR]
Yan-fang Ju1, Jin-ju Yang, Rong Liu
1Beijing Institute of Radiation Medicine, Beijing 100850, China. yanfangju@yahoo.com.cn
Aim:
To prepare monoclonal antibodies (mAbs) against Dicarbonyl/L-xylulose reductase (DCXR).
Methods:
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.
Results:
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.
Conclusion:
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.
