Generation and characterization of a polyclonal antibody against human high mobility group box 4

Yang,Fen; Li,Runsheng; Hong,Aizhen; Duan,Fei; Li,Yuhua

doi:10.3892/mmr.2013.1701

November 2013 Volume 8 Issue 5

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November 2013 Volume 8 Issue 5

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Article

Generation and characterization of a polyclonal antibody against human high mobility group box 4

Authors:
- Fen Yang
- Runsheng Li
- Aizhen Hong
- Fei Duan
- Yuhua Li
View Affiliations / Copyright

Affiliations: Shanghai Medical College, Fudan University, Shanghai 200032, P.R. China, Key Laboratory of Contraceptive Drugs and Devices of NPFPC, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, P.R. China
Pages: 1460-1464
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Published online on: September 24, 2013

https://doi.org/10.3892/mmr.2013.1701
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Abstract

A human high mobility group box 4 (hHMGB4) expression construct (pET‑28a/hHMGB4) was generated by cloning the hHMGB4 full‑length cDNA in the expression vector pET‑28a(+). The hHMGB4 fusion protein with His6‑Tag was prepared using E.coli BL21 (DE3) transformed with pET‑28a/hHMGB4 and purified via preparative SDS‑PAGE plus electroelution. Immunization of rabbits with the purified hHMGB4 generated polyclonal antibodies. The titer of the antiserum was determined to be 1:102,400 by ELISA analysis. Western blotting analysis showed that the antibody specifically recognized the recombinant hHMGB4 protein and also the endogenous hHMGB4 protein in prostate cancer cells. In addition, immunohistochemical staining analysis using the prepared antibody revealed marked hHMGB4 staining in the nuclei of the human prostate tissue. These data demonstrate that the anti‑hHMGB4 polyclonal antibody may be a useful reagent for the functional study of hHMGB4.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Generation and characterization of a polyclonal antibody against human high mobility group box 4

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Abstract

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