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[Preparation and application of monoclonal antibody against human vascular endothelial cadherin 5]
Jie Ding1, Yang He2, Jianfeng Yang3
1Clinical Laboratory, First Affiliated Hospital, Soochow University, Suzhou 215006, China.
Objective:
To prepare and identify monoclonal antibody (mAb) against human vascular endothelial cadherin 5 (VECDH5).
Methods:
BALB/c mice were immunized with recombinant VECDH5 protein for preparing mAb 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 mAb 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 mAb could specifically recognize and bind VECDH5. Epitope identification showed that the amino acid sequence recognized by 2C11 was LDREVVPWYNLTVEA.
Conclusion:
We have prepared mAb against human VECDH5, which has good binding ability and specificity.
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