Evaluation of primary opioid receptor antibodies for use in western blotting  H. Niwa, D.J. Rowbotham, D.G. Lambert  British Journal of Anaesthesia  Volume 108, Issue 3, Pages 530-532 (March 2012) DOI: 10.1093/bja/aes015 Copyright © 2012 The Author(s) Terms and Conditions

Fig 1 Western blotting images using non-specific KOP and MOP receptor antibodies. Samples were denatured and separated by SDS–PAGE, transferred to nitrocellulose and blotted with two antibodies for rMOP and with four for rKOP receptor. The optimized dilution factors used for antibodies are shown. The boxed region represents the expected molecular size of OP receptor protein. We used the CHO cells expressing rat KOP receptors (rKOP), rat MOP receptors (rMOP), human MOP receptors (hMOP), human KOP receptors (hKOP) as the positive control, wild-type of CHO cells lacking any opioid receptor expression; WT as the negative control. In a series of radioligand binding experiments, CHOrMOP, CHOrKOP, and CHOhMOP bound [3H]diprenorphine with Bmax and Kd of 266 fmol mg−1 protein and 0.19 nM, 231 fmol mg−1 protein and 0.15 nM, and 458 fmol mg−1 protein and 0.30 nM, respectively. British Journal of Anaesthesia 2012 108, 530-532DOI: (10.1093/bja/aes015) Copyright © 2012 The Author(s) Terms and Conditions