Preparation and characterization of a monoclonal antibody against the refolded and functional extracellular domain of rat P2X4 receptor

Tatsuhiro Igawa, Sadayuki Higashi, Yoshito Abe, Takatoshi Ohkuri, Hiroyuki Tanaka, Satoshi Morimoto, Tomohiro Yamashita, Tsuda Makoto, Kazuhide Inoue, Tadashi Ueda

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The P2X4 purinergic receptor is a key molecule in neuropathic pain, particularly in the allodynia after peripheral nerve injury. We therefore sought to establish an anti-P2X4 receptor monoclonal antibody that would be useful for detection and characterization of the P2X4 receptor. We first prepared the refolded extracellular domain of the rat P2X4 receptor expressed in Escherichia coli. Then, we established a B-cell hybridoma producing the monoclonal antibody for the head domain of the rat P2X4 receptor with strict recognition, including S-S bond formation. In addition, we succeeded in the detection and immune precipitation of rat P2X4 receptor molecules on cultured cells. As the antibody specifically binds to the rat P2X4 receptor molecule, it is expected that the established monoclonal antibody will be applicable as a tool for detecting increasing expression levels of the P2X4 receptor molecule accompanied with increasing intensity of neuropathic pain.

Original languageEnglish
Pages (from-to)275-282
Number of pages8
JournalJournal of Biochemistry
Volume153
Issue number3
DOIs
Publication statusPublished - Mar 1 2013

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Purinergic P2X4 Receptors
Rats
Monoclonal Antibodies
Molecules
Neuralgia
Cells
Peripheral Nerve Injuries
Hyperalgesia
Hybridomas
Immunoprecipitation
Escherichia coli
Cultured Cells
B-Lymphocytes
Head

All Science Journal Classification (ASJC) codes

  • Medicine(all)
  • Biochemistry
  • Molecular Biology

Cite this

Preparation and characterization of a monoclonal antibody against the refolded and functional extracellular domain of rat P2X4 receptor. / Igawa, Tatsuhiro; Higashi, Sadayuki; Abe, Yoshito; Ohkuri, Takatoshi; Tanaka, Hiroyuki; Morimoto, Satoshi; Yamashita, Tomohiro; Makoto, Tsuda; Inoue, Kazuhide; Ueda, Tadashi.

In: Journal of Biochemistry, Vol. 153, No. 3, 01.03.2013, p. 275-282.

Research output: Contribution to journalArticle

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AU - Tanaka, Hiroyuki

AU - Morimoto, Satoshi

AU - Yamashita, Tomohiro

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AU - Inoue, Kazuhide

AU - Ueda, Tadashi

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