[Preparation and identification of monoclonal antibody against UGP2].
The study was aimed to generate monoclonal antibodies (mAbs) against homo sapiens UDP-glucose pyrophosphorylase 2 (UGP2). Normal human liver tissues homogenized, and cytosolic proteins isolated by centrifugation were used to immunize BALB/c mice to generate mAbs by hybridoma technique. The mAbs were identified by ELISA, Western blot, and immunohistochemistry assay. The antibody specificity was confirmed by Uni-ZAP expression library screening. The results indicated that one hybridoma BAD062 secreting specific mAb against UGP2 was established. The Ig subclass of this mAb was IgG(2b) (kappa), and it could be used in ELISA, Western blot, immunohistochemistry assay. The antigen recognized by BAD062 mAb was localized in the hepatocyte cytoplasm, with molecular weight of 56 kD in the cytosolic proteins of human liver tissue. The BAD062 mAb was further confirmed by immunoscreening of Uni-ZAP XR liver cDNA expression library. It is concluded that a hybridoma cell line stably secretes specific mAb against UGP2. This mAb reacted with UGP2 in ELISA, Western blot, immunohistochemistry assay, and would be very useful for the UGP2 studies.
Wang
Wan
W
College of Life Science, Shandong Normal University, Jinan 250014, China.
Gao
Yuan
Y
Yang
Jin-Ju
JJ
Liu
Xiao-Lan
XL
Ju
Yan-Fang
YF
Liu
Li
L
Chen
Zhi-Cheng
ZC
Liu
Rong
R
Chi
Jun
J
Xing
Wei-Xian
WX
Gao
Jian-En
JE
An
Li-Guo
LG
Sun
Qi-Hong
QH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D000918:Antibody Specificity; D001483:Base Sequence; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000276:immunology*
D000911:Antibodies, Monoclonal; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase