1. [Preparation of rabbit polyclonal antibody against mouse ADAMTS2].
- Author
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Fu J, Miao S, and Wang L
- Subjects
- ADAM Proteins genetics, ADAM Proteins metabolism, ADAMTS Proteins, ADAMTS4 Protein, Animals, Antibody Specificity immunology, Blotting, Western, Enzyme-Linked Immunosorbent Assay, Female, Male, Mice, Inbred C57BL, Peptide Fragments genetics, Peptide Fragments immunology, Peptide Fragments metabolism, Procollagen N-Endopeptidase genetics, Procollagen N-Endopeptidase metabolism, Rabbits, Recombinant Proteins metabolism, ADAM Proteins immunology, Antibodies, Monoclonal immunology, Immune Sera immunology, Procollagen N-Endopeptidase immunology, Recombinant Proteins immunology
- Abstract
Objective: To obtain recombinant mouse collagenase ADAMTS2 (ADAM metallopeptidase with thrombospondin type 1 motif, 2) C terminal (1109-1213), and prepare the corresponding rabbit anti-ADAMTS2 polyclonal antibodies., Methods: The recombinant expression plasmid pGEX-6p-1-ADAMTS2 (1109-1213) was transformed into E.coli. The target protein was induced by IPTG and identified by mass spectrometry following affinity purification. The expressed and purified ADAMTS2 (1109-1213) protein was used to immunize New Zealand rabbits to prepare anti-ADAMTS2 polyclonal antibodies. The antibody titers were detected by ELISA and the antibody specificity by Western blotting., Results: The protein ADAMTS2 (1109-1213) was expressed in E.coli after IPTG induction, and with the purified protein, we prepared antiserum in the immunized rabbits. The titer of the antiserum reached over 1:160 000. The antiserum showed a good specificity., Conclusion: The high titer and specific rabbit anti-ADAMTS2 antibody has been prepared successfully.
- Published
- 2016