[Preparation and analysis of monoclonal antibody against EphA4 peptide].
OBJECTIVE
To prepare monoclonal antibody (MAb) against carboxy-terminus of E-phA4 and to analyze the immunological characteristics and significance of the antibody.


METHODS
Mice were immunized with a chemically synthesized peptide that had been conjugated to KLH (keyhole limpet hemocyamin). A panel of antibodies specifically against EphA4 were obtained by hybridoma technique. The immunological properties and significance of the MAb were analyzed by immunological and immunochemistry techniques.


RESULTS
A hybridoma cell line secreting anti-EphA4 MAb was established. The MAb reacted specifically with human, chicken and mouse EphA4, but did not cross-react with EphA5 and EphA7. The antibody could be used for immunochemical techniques such as ELISA, immunoprecipitation, Western blot and immunohistochemistry.


CONCLUSIONS
The anti-EphA4 MAb generated by immunizing a synthetic peptide possesses excellent immunological properties. MAb can be applied for immunological and immunohistochemical purpose and will become an important tool in the study of EphA4 and its ligand.
Yang
Jun-Jie
JJ
Departnent of Hematology, Second Xiangya Hospital, Central South University, Changsha 410011, China. zmyang126@hotmail.com
Weimin
Qi
Q
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology; D036122:Receptor, EphA4 / Q000276:immunology*
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D036122:Receptor, EphA4