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Preparation and application of monoclonal antibody against human vascular endothelial cadherin 5

DING Jie;HE Yang;YANG Jianfeng;ZUO Bin;YANG Binghua;Clinical Laboratory,First Affiliated Hospital,Soochow University;Jiangsu Provincial Institute of Hematology;Cyrus Tang Hematology Research Center,Soochow University;  
Objective To prepare and identify monoclonal antibody( m Ab) against human vascular endothelial cadherin 5( VECDH5). Methods BALB/c mice were immunized with recombinant VECDH5 protein for preparing m Ab using hybridoma technique. The positive clones were confirmed and selected by indirect ELISA for titer determination. Western blotting,flow cytometry,immunofluorescence staining, immunohistochemical staining were performed to identify the specificity and epitope. Results One hybridoma cell strain secreting specific m Ab against VECDH5 was obtained( 2C11). ELISA showed that the titer of the ascites was 1 ∶ 10 000. Western blotting,flow cytometry,immunofluorescence,immunohistochemistry demonstrated that the m Ab could specifically recognize and bind VECDH5. Epitope identification showed that the amino acid sequence recognized by 2C11 was LDREVVPWYNLTVEA. Conclusion We have prepared m Ab against human VECDH5,which has good binding ability and specificity.
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