2017

Edmund Wing Ong. Investigating the Effects of Prolonged Mu Opioid Receptor Activation Upon Opioid Receptor Heteromerization. A thesis submitted to the Graduate Program in Pharmacology & Toxicology in the Department of Biomedical and Molecular Sciences. Queen’s University Kingston, Ontario, Canada. March, 2017

...DRG neurons from adult rats were grown in culture for 4 days. They were then treated with either vehicle (saline) or morphine (10 μM, 48 hours) before fixation with 4% paraformaldehyde in 0.1 M phosphate buffer for 15 min at 37°C. Immunocytochemical labelling for μ and δ opioid receptors was accomplished with antibodies against the μ receptor (Neuromics) and the δ receptor...

Nathan Long, Bertha Long, Asma Mana, Dream Le, Lam Nguyen, Sima Chokr, Kevin Sinchak, Tamoxifen and ICI 182,780 activate hypothalamic G protein-coupled estrogen receptor 1 to rapidly facilitate lordosis in female rats. Hormones and Behavior. Available online 4 January 2017. http://dx.doi.org/10.1016/j.yhbeh.2016.12.013

... Briefly, every fourth MPN section was incubated in MOP primary antibody (1:5000; Neuromics Antibodies Edina, MN; Dewing et al., 2007 and Mills et al., 2004) and visualized using Fluorescein (FITC) staining associated with a Tyramide Signal Amplification kit (Perkin Elmer ... 


2016

Wen-Ling Dai, Feng Xiong, Bing Yan, Zheng-Yu Cao, Wen-Tao Liu, Ji-Hua Liu1, Bo-Yang Yu. Blockade of neuronal dopamine D2 receptor attenuates morphine tolerance in mice spinal cord. Scientific Reports 6, Article number: 38746 (2016). doi:10.1038/srep38746

...Immunofluorescence and Cell immunofluorescence

For immunofluorescence, mice were anesthetized and perfused with 0.01 M PBS followed by 4% paraformaldehyde on day 7. Lumbar spinal cords (L4-L6) were collected and post-fixed in the same 4% paraformaldehyde for one day and then cryo-protected in 30% sucrose for 3–5 days. The embedded blocks were sectioned as 25 μm thick58. Tissue sections from each group (4 mice, 8 images in each group) were blocked with 10% normal donkey serum and 0.3% Triton-X-100 (Sigma-Aldrich, St. Louis, MO, USA) for 2 h at room temperature. Then primary antibodies against mouse anti-D2DR (1:50, Santa Cruz Biotechnology, CA, USA), goat anti-IBA1 (1:200, Abcam, Cambridge, MA, USA), rabbit anti-GFAP (1:200, Abcam, Cambridge, MA, USA), or rabbit anti-NeuN (1:200, Abcam, Cambridge, MA, USA), rabbit anti-MOR (1:500, Neuromics, Edina, Minnesota, USA) were incubated overnight at 4 °C. Rabbit IgG (1:300; Jackson ImmunoResearch Laboratories, Inc., PA, USA) was used as an isotype control. Then the free-floating sections were washed with PBS, and incubated with the secondary antibody for 2 h. After washing out three times with PBS, the slices were incubated with DAPI (Southern Biotec, Birmingham, Alabama, USA).

For cell immunofluorescence, the cells in confocal dishes from each group (4 confocal dishes in each group) were washed with 0.01 M PBS and fixed in 4% paraformaldehyde at room temperature for 15 min, and then blocked with 10% normal donkey serum in 0.01 M PBS at room temperature for 2 h. Next, the cells were incubated with primary antibodies: rabbit anti-NeuN (1:200, Abcam, Cambridge, MA, USA), mouse anti-D2DR (1:50, Santa Cruz Biotechnology, CA, USA) and rabbit anti-MOR (1:500, Neuromics, Edina, Minnesota, USA) overnight at 4 °C. Rabbit IgG (1:300; Jackson ImmunoResearch Laboratories, Inc., PA, USA) was used as an isotype control. Then the cells in confocal dishes were washed with 0.01 M PBS and incubated with the secondary antibody for 2 h. After washing out three times with PBS, the cells were incubated with DAPI (Southern Biotec, Birmingham, Alabama, USA).

Confocal microscopy of dual antibody in both immunofluorescence and cell immunofluorescence were performed with a confocal laser-scanning microscope (Carl Zeiss LSM710, Germany). Images were randomly coded and transferred to a computer for further analysis. To obtain quantitative measurements, 8 images for each group were evaluated and photographed at the same exposure time to generate the raw data. Fluorescence intensities of the different groups were analyzed using Image Pro Plus 6.0 (Media Cybernetics, Silver Spring, MD, USA)....

Kazuya Hagimoto,Saki Takami,Fujio Murakami,Yasuto Tanabe. Distinct Migratory Behaviors of Striosome and Matrix Cells Underlying the Mosaic Formation in the Developing Striatum. The Journal of Comparative Neurology. 17 August 2016. DOI: 10.1002/cne.24096

... polyclonal rabbit anti-DARPP32 Ab (Cell Signaling Technology, Inc., Danvers, MA, USA)at 1:800, polyclonal guinea pig anti-μ-opioid receptor (MOR) Ab (Neuromics, Minneapolis,MN, USA) at 1:500, polyclonal rabbit anti-CTIP2 Ab (Abcam) at 1:500, ...

K. Lutfy, D. Parikh, D.L. Lee, Y. Liu, M.G. Ferrini, A. Hamid, T.C. Friedman. Prohormone convertase 2 (PC2) null mice have increased mu opioid receptor levels accompanied by enhanced morphine-induced antinociception, tolerance and dependence. Neuroscience. Available online 18 May 2016. doi:10.1016/j.neuroscience.2016.05.021

... were washed in PBS, quenched in 3% hydrogen peroxide for endogenous peroxidase activity,washed in PBS, blocked in PBS containing 10% normal goat serum and 0.15% Triton X-100, and incubated with the MOP primary polyclonal antibody (1:500) (Neuromics, Edina, MN ...

Wendy M. Walwyn, Wenling Chen, Hyeyoung Kim, Ani Minasyan, Helena S. Ennes, James A. McRoberts, and Juan Carlos G. Marvizón. Sustained Suppression of Hyperalgesia during Latent Sensitization by μ-, δ-, and κ-opioid receptors and α2A Adrenergic Receptors: Role of Constitutive Activity. The Journal of Neuroscience, 6 January 2016, 36(1): 204-221; doi: 10.1523/JNEUROSCI.1751-15.2016

...probed with antibodies to MOR (RA10104; Neuromics), p-Ser375-MOR (pMOR, RA18001; Neuromics) and p-Tyr416-Src family kinase (pSFK...serine-phosphorylated MOR (p-Ser375-MOR, RA18001; Neuromics) and a different MOR antibody (RA10104...

2015

Jan Fraessdorf, Markus W. Hollmann , Iris Hanschmann, André Heinen, Nina C. Weber, Benedikt Preckel, Ragnar Huhn. Role of Endogenous Opioid System in Ischemic-Induced Late Preconditioning. Published: July 30, 2015DOI: 10.1371/journal.pone.0134283.

...Antibodies for μ-OR and δ-OR were purchased from Neuromics (Minneapolis, USA)...

Ying-Biao Chen, Fen-Sheng Huang, Ban Fen, Jun-Bin Yin, Wei Wang and Yun-Qing L nhibitory effects of endomorphin-2 on excitatory synaptic transmission and the neuronal excitability of sacral parasympathetic preganglionic neurons in young rat. Journal Name: Frontiers in Cellular Neuroscience, ISSN: 1662-5102, Article type: Original Research Article,Received on: 02 Apr 2015, Accepted on: 12 May 2015, Provisional PDF published on: 12 May 2015.

... guinea pig antiserum against MOR (1:1000, GP10106, Neuromics, Edina, Minnesota, USA)...

Sherika N. Smith, Candler Paige, Kandy T. Velazquez, Terika P. Smith, Srinivasa N. Raja, Steven P. Wilson, Sarah M. Sweitzer. Injury-specific promoters enhance herpes simplex virus mediated gene therapy for treating neuropathic pain in rodents. doi:10.1016/j.jpain.2014.12.007

...: MOR (Neuromics, RA10104, 1:1000)...

2014

Joanna Mika, Katarzyna Popiolek-Barczyk, Ewelina Rojewska, Wioletta Makuch, Katarzyna Starowicz, Barbara Przewlocka. Delta-Opioid Receptor Analgesia Is Independent of Microglial Activation in a Rat Model of Neuropathic Pain. Published: August 08, 2014DOI: 10.1371/journal.pone.0104420.

... rabbit anti-MOR, 1:400, rabbit anti-KOR, 1:400 and rabbit anti-DOR, 1:400, Neuromics...

Xue-Long Zhou, Li-Na Yu1, Yin Wang, Li-Hui Tang, Yu-Nan Peng, Jun-Li Cao and Min Yan. Increased methylation of the MOR gene proximal promoter in primary sensory neurons plays a crucial role in the decreased analgesic effect of opioids in neuropathic pain. Zhou et al. Molecular Pain 2014, 10:51.http://www.molecularpain.com/content/10/1/51

Western Blot: Tissue samples were homogenized in lysis buffer (12.5 μl/mg tissue) containing a mixture of protease inhibitors (Roche) and PMSF (Sigma). After incubating in ice for 30 min, samples were centrifuged at 10,000 rpm for 15 min at 4°C. The supernatants were used for Western blotting. Equal amount of protein (~30 μg) was loaded and separated in 10% Tris-Tricine SDS-PAGE gel. The resolved proteins were transferred onto polyvinilidene difluoride (PVDF) membranes (Amersham Bioscience). The membranes were blocked in 5% non-fat milk for 1 h at room temperature (RT), and incubated overnight at 4°C with rabbit anti-MOR antibody (Neuromics, 1:1,000) or anti-GAPDH antibody (Cell Signaling Technology, 1:2000) primary antibody. The blots were then incubated with the secondary antibody, goat anti-rabbit IgG conjugated with horseradish peroxidase (HRP) (1:1000, Cell Signaling Technology), for 2 h at room temperature. Signals were finally visualized using enhanced chemiluminescence (ECL, Pierce) and the blots were exposed by The ChemiDoc™ XRS + image system (Bio-Rad; Hercules).

Immunnohistochemistry: Mice were deeply anesthetized and undergone sternotomy, intracardially perfuse with 20 ml saline followed by 4% ice-cold paraformaldehyde in 0.1 mol/l phosphate buffer (PB). L4-L5 DRGs, L4-L5 spinal cord segments, sciatic nerves, and plantar surface of hindpaw skin were removed, post-fixed in 4% paraformaldehyde for 3 h, and subsequently allowed to equilibrate in 30% sucrose in PB overnight at 4°C. DRG and transverse spinal cord sections (15 μm), sciatic nerve sections (10 μm) and skin sections (40 μm) were cut on a cryostat and every fifth section was collected in PB. After washing in phosphate buffer saline (PBS), the tissue sections were incubated in PBS containing 5% normal goat serum and 0.3% TritonX-100 at room temperature for 30 min. The sections were incubated overnight at 4°C with rabbit antiMOR antibody (Neuromics, 1:500) followed by Rhodamine Red™-X goat anti-rabbit IgG (Invitrogen, 1:500) for 2 h at room temperature. Nonspecific staining was determined by excluding the primary antibodies. Sections were rinsed, mounted, and cover-slipped with glycerol containing 2.5% of anti-fading agent DABCO (Sigma) and stored at −20°C in the dark. Images were captured using a fluorescent microscope (DMIRB, Leica, Germany).

Fiona B Carr, Sandrine M Géranton and Stephen P Hunt. Descending controls modulate inflammatory joint pain and regulate CXC chemokine and iNOS expression in the dorsal horn. Molecular Pain 2014, 10:39 doi:10.1186/1744-8069-10-39. Published: 20 June 2014.

...mu opioid receptor, 1:10,000, Neuromics (RA10104)...

Charlie H.T. Kwok, Ian M. Devonshire, Andrew J. Bennett, Gareth J. Hathway. Postnatal maturation of endogenous opioid systems within the periaqueductal grey and spinal dorsal horn of the rat. PAIN - January 2014 (Vol. 155, Issue 1, Pages 168-178, DOI: 10.1016/j.pain.2013.09.0220.

...rabbit anti-MOR (Neuromics, Edina, MN, USA; 1:1000 with tyramide signal amplification protocol)...

J. Desroches, J.-F. Bouchard, L. Gendron, P. Beaulieu. Involvement of cannabinoid receptors in peripheral and spinal morphine analgesia ☆ Neuroscience, Volume 261, 7 March 2014, Pages 23–42. http://dx.doi.org/10.1016/j.neuroscience.2013.12.030.

...The floating sections were then incubated in 1% sodium borohydride in PBS for 30 min, rinsed twice with PBS, and incubated for 30 min at room temperature in a blocking solution containing 3% normal goat serum (NGS) and 0.3% Triton X-100 in PBS. The sections were then incubated overnight at 4 °C with the guinea pig anti-MOP primary antibody (cat# GP10106; Neuromics, Minneapolis, MN, USA) diluted 1:1000 in the blocking solution. The floating sections were then washed in PBS and incubated with a goat anti-guinea pig secondary antibody conjugated with Alexa Fluor 488 (Molecular Probes, Invitrogen, Carlsbad, CA, USA) at a concentration of 1:1000 in PBS for 2 h at room temperature...

2013

Charlie H.T. Kwok, Ian M. Devonshire, Andrew J. Bennett, Gareth J. Hathway Postnatal maturation of endogenous opioid systems within the periaqueductal grey and spinal dorsal horn of the rat. PAIN®, Available online 27 September 2013. http://dx.doi.org/10.1016/j.pain.2013.09.022. http://www.sciencedirect.com/science/article/pii/S0304395913005198.

...rabbit anti-MOR (Neuromics, Edina, MN, USA; 1:1000 with tyramide signal amplification protocol)...

...Shao-Qiu Hea, Fei Yanga, Federico M. Perezb, Qian Xua, Ronen Shechtera, Yong-Kwan Cheong, Alene F. Carteret, Xinzhong Dong, Sarah M. Sweitzer, Srinivasa N. Raja, 1,Yun Guan. Tolerance to the anti-allodynic effects of the peripherally acting opioid, loperamide hydrochloride, develops in nerve-injured rats. doi.org/10.1016/j.pain.2013.07.023.

... MOR (1:2,000, Neuromics), phospho-MOR (p-MOR-Ser375; 1:1,000, Neuromics)...

2011-2012

Shafaq Sikandar, Kirsty Bannister, Anthony H. Dickenson. Brainstem facilitations and descending serotonergic controls contribute to visceral nociception but not pregabalin analgesia in rats. http://dx.doi.org/10.1016/j.neulet.2012.05.009

...For MOR staining, a rabbit polycolonal μ-opioid receptor primary antibody (1:10,000 TTBS; Neuromics, MN, USA, RA10104) was used with tyramide signal amplification (TSA, Perkin Elmer, USA)...

Andrew C. Eschenroeder, Allison A. Vestal-Laborde, Emilse S. Sanchez, Susan E. Robinson, Carmen Sato-Bigbee. Oligodendrocyte responses to buprenorphine uncover novel and opposing roles of μ-opioid- and nociceptin/orphanin FQ receptors in cell development: Implications for drug addiction treatment during pregnancy. Glia Volume 60, Issue 1, pages 125–136, January 2012.

...The MOR and NOP receptor antibodies were from Neuromics (Edina, MN) and used for immunocytochemistry (1:100)and western blotting (1:500)...

De Felice, Milena, Sanoja, Raul, Wang, Ruizhong, Vera-Portocarrero, Louis, Oyarzo, Janice, King, Tamara, Ossipov, Michael H., Porreca, Frank. Engagement of descending inhibition from the rostral ventromedial medulla protects against chronic neuropathic pain. PAIN, 152 (12), p.2701-2709, Dec 2011.

...MOR (rabbit, 1:2000, Neuromics, Minneapolis, MN), IB4 (Ab to lectin; 1:1000, Vector Laboratories, Burlingame, CA), P2X3 (rabbit, 1:5000, Neuromics)...

Groer, Chad E, Cullen L. Schmid, Alex M. Jaeger and Laura M. Bohn. AGONIST-DIRECTED INTERACTIONS WITH SPECIFIC βARRESTINS DETERMINE MU OPIOID RECEPTOR TRAFFICKING, UBIQUITINATION, AND DEPHOSPHORYALTION. First Published on July 14, 2011, doi: 10.1074/jbc.M111.248310.

....MOR Phosphorylation- Western Blotting.   HA-MOR WT and βarr1/2 KO MEFs were serum  starved for 15 minutes, followed by a 10 minute treatment with vehicle (PBS), DAMGO (1 µM), or  morphine (10 µM).  The HA-MOR was  immunoprecipitated from cell lysates and analyzed  by western blot using antibodies against the MOR  phosphorylated at Serine 375 (Ser375; Cell
Signaling, Danvers, MA; 1:500) and the Cterminus of the MOR (Neuromics, Edina, MN;   1:1000).  Densitometry was assessed on the prominent ~75 kDa band using Kodak imaging  software, and the level of phosphorylated MOR  detected was normalized to the amount of total  MOR immunoprecipitated in each sample...  

Shao-Qiu He, Zhen-Ning Zhang, Ji-Song Guan, Hong-Rui Liu, Bo Zhao, Hai-Bo Wang, Qian Li, Hong Yang. Facilitation of m-Opioid Receptor Activity by Preventing d-Opioid Receptor- Mediated Codegradation. DOI 10.1016/j.neuron.2010.12.001.

...guinea pig anti-MOR antibody (1:400,Neuromics)...

2010-2008

...guinea pig anti-muOR (1:1000; Neuromics)...

Ryan J. Horvathab, Alfonso Romero-Sandoval, Joyce A. De Leo. Inhibition of microglial P2X4 receptors attenuates morphine tolerance, Iba1, GFAP and μ opioid receptor protein expression while enhancing perivascular microglial ED2. PAIN®. Pages 401-413 (September 2010).

...μ opioid receptor (1:5000, Neuromics), CGRP (1:500, Neuromics)...

Ying He, Cheng Yang, Chelsea M. Kirkmire, and Zaijie Jim Wang. Regulation of Opioid Tolerance by let-7 Family MicroRNA Targeting the µ Opioid Receptor. The Journal of Neuroscience, July 28, 2010, 30(30):10251-10258; doi:10.1523/JNEUROSCI.2419-10.2010. 

Hai-Bo Wanga, Bo Zhaoa, Yan-Qing Zhonga, Kai-Cheng Li, Zi-Yan Li, Qiong Wang, Yin-Jing Lua, Zhen-Ning Zhang, Shao-Qiu He, Han-Cheng Zheng, Sheng-Xi Wu, Tomas G. M. Hökfelt, Lan Baob, and Xu Zhanga. Coexpression of δ- and μ-opioid receptors in nociceptive sensory neurons. PNAS July 20, 2010 vol. 107 no. 29 13117-13122.

 Immunostaining. Adult rats, mice, and Oprd1 exon 1-deleted mice were fixed. Cryostat sections of L4 and L5 DRGs and spinal cord segments were processed for immunofluorescence staining (13) with Rb anti-DOR13–17 (1:2,000–1:60,000; DiaSorin and 1:4,000–1:60,000; Neuromics), Rb anti-DOR12–18 (1:30,000–1:120,000; Alomone), Rb anti-DOR1358–372 (1:1,000–1:2,000; Lifespan Biosciences), Rb anti-MOR (1:1,000; Neuromics); guinea pig anti-SP (1:500; Neuromics), and mouse anti-CGRP (1:1,000; Biogenesis) antibodies. IB4-labeling was carried out with fluorescein-labeled GSL I-IB4 (1:200). The Myc-DOR1–transfected HEK293 cells and neurons were fixed and processed with mouse anti-Myc antibodies (1:500; DSHB). Nuclear DAPI staining was used to indicate HEK293 cells in control experiments.

 Immunoblotting.The samples were processed for SDS/PAGE, transferred, probed with Rb antibodies against MOR (1:500; Neuromics), phospho-DOR1 (1:1,000; Neuromics), phospho-MOR (1:1,000; Neuromics), Myc (1:500; DSHB), Flag (1:1,000; Sigma), or actin (1:50,000; Chemicon) and visualized with enhanced chemiluminescence (19).

 .... Antibodies for δOR (mouse; Neuromics; RA10101)...

Dominika Labuz, Yvonne Schmidt, Anja Schreiter, Heike L. Rittner, Shaaban A. Mousa and Halina Machelska. Immune cell–derived opioids protect against neuropathic pain in mice. J. Clin. Invest. doi:10.1172/JCI36246. Copyright © 2009, The American Society for Clinical Investigation

κ-opioid receptor (1:800, Neuromics)

...For MOR internalization, rabbit primary antibodies directed against MOR (1:24,000; Neuromics, Minneapolis, MN) were used. Sections processed for fluorescence were incubated in blocking buffer (Tyramide Signal Amplification kit, NEN Life Science Products, Boston, MA), and then in biotin conjugated goat anti-rabbit IgG (Vector Laboratories)...

Marta Pinto, Marta Sousa, Deolinda Lima, Isaura Tavares. Participation of mu-opioid, GABAB, and NK1 receptors of major pain control medullary areas in pathways targeting the rat spinal cord: Implications for descending modulation of nociceptive transmission. Volume510 Issue 2 Pages 175 - 187 Published Online: 9 Jul 2008. Copyright © 2008 Wiley-Liss, Inc., A Wiley Company.

...The antibody against MOR (Neuromics; Cat No. RA10104, Lot No 400058)...

Sasaki, Atsushi a; Nakashima, Yasutaka a; Takasaki, Ichiro c; Andoh, Tsugunobu a; Shiraki, Kimiyasu b; Kuraishi, Yasushi a d. Morphine inhibits herpetic allodynia through [mu]-opioid receptors induced in A[beta]-fiber neurons. Neuroreport. 19(9):975-979, June 11, 2008

...The dorsal root ganglion (DRG) at the L5 level was removed; transcardial perfusion, postfixation, and cryoprotection were done as described previously [13]. They were cut on a freezing microtome (Leica CM 3050S IV, Nussloch, Germany) at a 16-mm thickness. After being treated with a blocking solution (1.5% fetal bovine serum and 0.2% Triton X-100 in the phosphate-buffered saline) for 30 min at room temperature, sections were incubated with rabbit anti-MOR antibody (1 : 500; Neuromics Inc., Minneapolis, Minnesota, USA) at 41oC for 2 days. After washing, the sections were reacted with Alexa 488-conjugated donkey anti-rabbit immunoglobulin G antibody (1 : 1000; Molecular Probes Inc., Eugene, Oregon, USA) for 2 h at room temperature.Fluorescence signals were observed using Radiance 2000 confocal system (Bio-Rad Microscopy Division, Cambridge, Massachusetts, USA)...

Zhang, Guohua M.D., Ph.D.; Mohammad, Husam; Peper, Brad D.; Raja, Srinivasa M.D; Wilson, Steven P. Ph.D; Sweitzer, Sarah M. Ph.D.  Enhanced Peripheral Analgesia Using Virally Mediated Gene Transfer of the [mu]-Opioid Receptor in Mice.
Anesthesiology - Volume 108, Issue 2 (February 2008)  -  Copyright © 2008 American Society of Anesthesiologists, Inc.

...Blocking was followed by a 24-h incubation at 4°C in polyclonal rabbit anti-mu-OR (1:1,000; Neuromics, Minneapolis, MN)...

2007-2003

...Sections required for electron microscopy were permeabilized in 50% ethanol for 30 min at room temperature, washed in PBS then incubated in a H2O2 solution for 30 min to block endogenous peroxidase activity. Sections for light microscopy were not treated with 50% ethanol for permeabilization, but had 0.1% TritonX100 added to the primary antibody solution. Tissue processing for light and electron microscope viewing has been described in detail previously (Deuchars et al. 2001a). Briefly, sections were incubated in polyclonal antiserum corresponding to residues 1359–1403 of the carboxy-terminus of the rat mu-opioid receptor (Neuromics, Minneapolis, USA), 1:2000 in PBS. This was localized with secondary biotinylated donkey anti-rabbit (1:200), prior to detection with extra-avidin peroxidase (1:1500) and visualization with DAB. Sections required for light microscopy were dehydrated in increasing concentrations of ethanol and mounted in DPX. Sections for electron microscopy were postfixed in 0.5% osmium tetra oxide and dehydrated in graded ethanol concentrations, 50%, 70%, 90% and 100%. Sections were immersed in Durcupan ACM resin for 12–18 h, mounted on slides, cover-slipped then placed in the oven at 60°C for 48 h to polymerize the resin. Slices were viewed at the light and electron microscope level...

Phoebe Dewing, Marissa I. Boulware,Kevin Sinchak, Amy Christensen, Paul G. Mermelstein,  and Paul Micevych. Membrane Estrogen Receptor-{alpha} Interactions with Metabotropic Glutamate Receptor 1a Modulate Female Sexual Receptivity in Rats. doi:10.1523/JNEUROSCI.0592-07.2007.

....... rabbit primary antibodies directed against MOR (1:24000; Neuromics, Minneapolis) ...

S.-R. Chen, A. Prunean, H.-M. Pan, K.L. Welker and H.-L. Pan. Resistance to morphine analgesic tolerance in rats with deleted transient receptor potential vanilloid type 1-expressing sensory neurons. doi:10.1016/j.neuroscience.2006.12.016

.... antibodies (rabbit anti-μ opioid receptors, dilution 1:1000, Neuromics; and guinea-pig anti-TRPV1, ...

Eunhae Kim, Amy L. Clark , Alexi Kiss, Jason W. Hahn, Robin Wesselschmidt. MU AND KAPPA OPIOIDS INDUCE THE DIFFERENTIATION OF EMBRYONIC STEM CELLS TO NEURAL PROGENITORS JBC Papers in Press. Published on September 1, 2006 as Manuscript M603862200

Thomas F. Finnegan, Shao-Rui Chen, and Hui-Lin Pan. Effect of the µ Opioid on Excitatory and Inhibitory Synaptic Inputs to Periaqueductal Gray-Projecting Neurons in the Amygdala. JPET 312:441-448, 2005.

...The second primary antibody, rabbit anti-µ receptor (1:2000; Neuromics, Northfield, MN), was then applied for 2 h at room temperature and 24 h at 4°C. Sections were rinsed in Tris-buffered saline and 0.05% Tween 20 and exposed to the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (1:100; Jackson ImmunoResearch Laboratories Inc.) for 90 min at room temperature...

Byron C. Yoburn*, Vishal Purohit, Kaushal Patel, Qiuyu Zhang. Opioid agonist and antagonist treatment differentially regulates immunoreactive Mu-opioid receptors and dynamin-2 in vivo. European Journal of Pharmacology 498 (2004) 87–96

...Primary Antibody (Mu-opioid receptor, 1:500; Neuromics, Northfield, ... Western blot protocol...

Tara A. Macey, Janet D. Lowe, and Charles Chavkin. Mu Opioid Receptor Activation of ERK1/2 Is GRK3 and Arrestin Dependent in Striatal Neurons. Published online 2006 September 18. doi: 10.1074/jbc.M60427820. 

..guinea pig anti-mu opioid receptor (1/1000 dilution, Neuromics, Minneapolis, MN)... 

Shuxing Wang, Grewo Lim, Liling Yang, Qing Zeng, Backil Sung, J.A. Jeevendra Martyn and Jianren Mao.

A rat model of unilateral hindpaw burn injury: Slowly developing rightwards shift of the morphine dose–response curve. doi:10.1016/j.pain.2005..03.044.

...MOR (1:2000; Rabbit anti-rat MOR, Neuromics Inc.)...

Wu Z, Chen S and Pan H. Differential Sensitivity of N- and P/Q-Type Ca2+ Channel Currents to a µ Opioid in Isolectin B -Positive and -Negative Dorsal Root Ganglion Neurons. JPET 311:939-947, 2004 

...Sections were incubated with the primary antibody (rabbit anti-µ opioid receptor, dilution 1:1000; Neuromics... 

Truong W, Cheng C, Xu Q, Li X, and Zochodne, D. Mu Opioid receptors and analgesia at the site of a peripheral nerve injury Annals of Neurology Volume 53, Issue 3 , Pages 366 - 375

Chen S and Pan H. Antinociceptive Effect of Morphine, but not Mu Opioid Receptor Number, Is Attenuated in the Spinal Cord of Diabetic Rats. Anesthesiology 2003; 99:1409–14. 

...sections were then incubated with the primary antibody (rabbit antibody to mu opioid receptor; dilution, 1:1,000; Neuromics... 

Jones T, Sweitzer S, Wilson S, and Yeomans D. (2003). Afferent fiber-selective shift in opiate potency following targeted opioid receptor knockdown. Pain 106 (2003) 365–371 

...antisera solution (1:1000) against the mOR (Neuromics... 

Western blot analysis. Western blot analysis was performed as previously described (Tang et al., 2006) using the anti-µ opioid receptor antibody (1:1000; Neuromics). The expression of β-actin was similarly determined from the same blots using a monoclonal antibody (1:10,000; Sigma).

For immunofluorescence analysis, the antibody for hDcp1a (Santa Cruz Biotechnology) and MOR were used at 1:500 and 1:5000 dilutions, respectively. Secondary anti-goat and anti-mouse antibodies labeled with Alexa 488 and Alexa 594 fluorochromes (Invitrogen), respectively, were used at 1:500 dilutions.