[Preparation and characterization of monoclonal antibodies against RalB].
OBJECTIVE
In order to explore the expression of RalB (ras related; GTP binding protein B) in mammal eucaryotic cell, we prepared and characterized monoclonal antibodies against RalB.


METHODS
Hybridomas were generated by the fusion with Sp2/0 myelomas and spleen cells, which were from mice immunized with RalB recombinant proteins. The monoclonal antibodies against RalB were then used to identify the expression of RalB in mammal eucaryotic cell, including normal hepatic cell and hepatoma carcinoma cells, by Western blot and Immunohistochemistry.


RESULTS
Two hybridoma cell lines, F001, F002, had been produced, each of which stably secrets antibodies against RalB. Subclass of IgG are both belonged to IgG1. Immunohistochemistry demonstrated that RalB was presented in plasma membrane of hepatoma tissue. Western-blot showed that RalB was expressed in all concerned cell.


CONCLUSIONS
The monoclonal antibodies against RalB protein have been successfully prepared, which should provide useful reagent for further investigation into the biological function of RalB.
Pan
Hua-zheng
HZ
School of Biological Technology, Southern Medical University, Guangzhou 510515, China.
Huang
Xiang
X
Yao
Xiao-yan
XY
Wang
Sui-hai
SH
Du
Hong-yan
HY
Ji
Chao-neng
CN
Li
Ming
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005260:Female; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D051841:Transcription Factor RelB / Q000276:immunology*; D020725:ral GTP-Binding Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; C497594:Relb protein, mouse; D051841:Transcription Factor RelB; D020725:ral GTP-Binding Proteins