[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.

CONCLUSION

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.