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Quantitative Detection of µ Opioid Receptor: Western Blot Analyses Using µ Opioid Receptor Knockout Mice

Shinya Kasai1, Hideko Yamamoto, Etsuko Kamegaya

  • 1Division of Psychobiology, Tokyo Institute of Psychiatry, Tokyo 156-8585, Japan.

Current Neuropharmacology
|September 3, 2011
PubMed

Insights

A new antibody (N38) accurately detects the molecular weight of the mu opioid receptor (MOP) in mouse brains. This tool shows promise for quantifying MOP expression in substance dependence research.

Area of Science:

  • Neuroscience
  • Pharmacology

Background:

  • Mu opioid receptor (MOP) expression changes are linked to substance dependence.
  • Previous anti-MOP antibodies yielded inconsistent molecular weight (MW) data.

Purpose of the Study:

  • To generate and validate a new anti-MOP antibody (N38) for accurate MOP detection.
  • To determine the intrinsic MW of MOP using Western blot analysis.

Main Methods:

  • Generated a novel anti-MOP antibody (N38) targeting the N-terminus (amino acids 1-38).
  • Performed Western blot analysis on wildtype and MOP knockout mouse brain lysates.
  • Isolated and analyzed plasma membrane fractions.

Main Results:

  • The N38 antibody detected MOP bands at 60-67 kDa in wildtype brain plasma membranes.
  • No bands were detected in MOP knockout brain samples, confirming specificity.
  • Detected bands showed a semi-linear density response to protein concentration (3-30 µg).

Conclusions:

  • The N38 antibody specifically detects intrinsic MOP at a consistent MW range.
  • This antibody shows potential for quantitative MOP expression analysis in neuroscience research.
Keywords:
Knockout miceWestern blot analysis.quantificationµ opioid receptor

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