[Preparation of rabbit polyclonal antibody against mouse ADAMTS2]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Mar;32(3):393-6.
[Article in Chinese]

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.

MeSH terms

  • ADAM Proteins / genetics
  • ADAM Proteins / immunology*
  • ADAM Proteins / metabolism
  • ADAMTS Proteins
  • ADAMTS4 Protein
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity / immunology
  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Immune Sera / immunology*
  • Male
  • Mice, Inbred C57BL
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Peptide Fragments / metabolism
  • Procollagen N-Endopeptidase / genetics
  • Procollagen N-Endopeptidase / immunology*
  • Procollagen N-Endopeptidase / metabolism
  • Rabbits
  • Recombinant Proteins / immunology*
  • Recombinant Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Immune Sera
  • Peptide Fragments
  • Recombinant Proteins
  • ADAM Proteins
  • ADAMTS Proteins
  • Adamts2 protein, mouse
  • Procollagen N-Endopeptidase
  • ADAMTS4 Protein