Archives of Pharmacal Research

, Volume 18, Issue 2, pp 113–117 | Cite as

Preparation of δ-opioid receptor-specific antibodies using molecular cloned genes

  • Ae-Young Kim
  • Shee-Yong Lee
  • Kyeong-Man Kim
Research Articles
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Abstract

We re-cloned mouse δ-opioid receptor from NG108-15 cells using RT-PCR, and confirmed it by restriction analysis and by sequencing the beginning and end part of the amplified DNA. When transiently expressed in COS-7 cells, cloned δ-opioid receptor showed saturable and specific binding to [3H]naloxone with very similar binding parameters to originally reported ones. To make antibodies specific for the δ-opioid receptor, the carboxy tail of the δ-opioid receptor, which is unique to the δ-opioid receptor compared with other opioid receptors, was expressed in bacteria as a fusion proteinwith glutathione S-transferase. Purified fusion protein was injected into rabbits to produce antibodies. The antibodies produced were highly selective for δ-opioid receptor when tested by western blotting using membrane proteins prepared from transfected COS-7 cells. Cloned δ-opioid receptor and antibodies specific for δ-opioid receptor are going to be valuable tools for studying pharmacological actions of the δ-opioid receptor and morphine dependence.

Key words

δ-Opioid receptor Antibodies Cloning Binding study 
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Copyright information

© The Pharmaceutical Society of Korea 1995

Authors and Affiliations

  • Ae-Young Kim
    • 1
  • Shee-Yong Lee
    • 1
  • Kyeong-Man Kim
    • 1
  1. 1.Pharmacology Laboratory, College of PharmacyChonnam National UniversityKwang-JuKorea

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