[Preparation and characterization of monoclonal antibody against DCXR].
OBJECTIVE
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.


CONCLUSIONS
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.
Ju
Yan-fang
YF
Beijing Institute of Radiation Medicine, Beijing 100850, China. yanfangju@yahoo.com.cn
Yang
Jin-ju
JJ
Liu
Rong
R
Gao
Yuan
Y
Liu
Xiao-lan
XL
Gao
Jian-en
JE
Sun
Qi-hong
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000917:Antibody Formation; D000918:Antibody Specificity; D015153:Blotting, Western; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013401:Sugar Alcohol Dehydrogenases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D013401:Sugar Alcohol Dehydrogenases; C023300:L-xylulose reductase