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Volume 9 Issue 4
Dec.  2018
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Article Navigation >  Journal of Tropical Biology  > 2018  >  9(4): 370-376

ZHANG Shengnan, WU Ying, SUN Yun, Cao Zhenjie, ZHOU Yongcan. Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM[J]. Journal of Tropical Biology, 2018, 9(4): 370-376. doi: 10.15886/j.cnki.rdswxb.2018.04.002
Citation: ZHANG Shengnan, WU Ying, SUN Yun, Cao Zhenjie, ZHOU Yongcan. Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM[J]. Journal of Tropical Biology, 2018, 9(4): 370-376. doi: 10.15886/j.cnki.rdswxb.2018.04.002
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Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM

doi: 10.15886/j.cnki.rdswxb.2018.04.002

  • Ocean College, Hainan University/Hainan UniversityHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology/State Key Laboratory of Marine Resource Utilization in South China Sea/, Haikou, Hainan 570228

  • Received Date: 2018-09-10
  • Rev Recd Date: 2018-10-18
  • Immunoglobulin M（ IgM） was isolated from the serum of Trachinotus ovatusby using ammonium sulfate fractionation and then purified by using Protein A-sepharose affinity chromatography. Furthermore,the purified IgM product was used to prepare a polyclonal antibody. In addition,the titer and specificity of the polyclonal antibody were analyzed by the ELISA and the Western blot,respectively. The results showed that the ammonium sulfate fractionation precipitated most proteins from the sera of T. ovatus,but the IgM proteins with heavy and light chains were crude with some heteroproteins. Furthermore,the IgM product purified by the Protein A-sepharose affinity chromatography possessed two obvious bands（ heavy chain and light chain）. Moreover,the prepared anti-sera had a high titer of 1 ∶ 25 600 when assayed by the ELISA. The Western blot analysis showed that the IgM polyclonal antibody of T. ovatus contained the target bands,indicating the IgM polyclonal antibody was prepared successfully and could be used in correlation analysis in the coming future.
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Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM

doi: 10.15886/j.cnki.rdswxb.2018.04.002
  • Ocean College, Hainan University/Hainan UniversityHainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology/State Key Laboratory of Marine Resource Utilization in South China Sea/, Haikou, Hainan 570228
  • Received Date: 2018-09-10
  • Rev Recd Date: 2018-10-18

Abstract: Immunoglobulin M（ IgM） was isolated from the serum of Trachinotus ovatusby using ammonium sulfate fractionation and then purified by using Protein A-sepharose affinity chromatography. Furthermore,the purified IgM product was used to prepare a polyclonal antibody. In addition,the titer and specificity of the polyclonal antibody were analyzed by the ELISA and the Western blot,respectively. The results showed that the ammonium sulfate fractionation precipitated most proteins from the sera of T. ovatus,but the IgM proteins with heavy and light chains were crude with some heteroproteins. Furthermore,the IgM product purified by the Protein A-sepharose affinity chromatography possessed two obvious bands（ heavy chain and light chain）. Moreover,the prepared anti-sera had a high titer of 1 ∶ 25 600 when assayed by the ELISA. The Western blot analysis showed that the IgM polyclonal antibody of T. ovatus contained the target bands,indicating the IgM polyclonal antibody was prepared successfully and could be used in correlation analysis in the coming future.

ZHANG Shengnan, WU Ying, SUN Yun, Cao Zhenjie, ZHOU Yongcan. Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM[J]. Journal of Tropical Biology, 2018, 9(4): 370-376. doi: 10.15886/j.cnki.rdswxb.2018.04.002
Citation: ZHANG Shengnan, WU Ying, SUN Yun, Cao Zhenjie, ZHOU Yongcan. Purification of Serum IgM from Trachinotus ovatus and Preparation of Mouse Sera Anti-IgM[J]. Journal of Tropical Biology, 2018, 9(4): 370-376. doi: 10.15886/j.cnki.rdswxb.2018.04.002
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