Xing J. & Li J. Bradykinin Contributes to Sympathetic and Pressor Responses Evoked by Activation of Skeletal Muscle Afferents P2X in Heart Failure. Cell Physiol Biochem 2016;39:2101-2109. DOI:10.1159/000447906

...Western Blotting Analysis

The L4-L6 DRGs from both sides were removed after the rats were anesthetized by overdose of isoflurane followed by cervical dislocation and decapitation. L4-L6 levels of DRGs were selected since they contain cell bodies of sensory afferents innervating the hindlimbs. Samples were obtained from individual animals. Briefly, the concentration of protein in the homogenate was determined using a BCA reagent after the tissues were lyzed and centrifuged. Kinin B2 and P2X3 proteins were loaded onto gel, respectively. After electrophoresis, the proteins were electrotransferred onto polyvinylidene difluoride membranes. The membranes were then incubated with the primary antibodies: guinea pig anti-B2 (1:500; Santa Cruz) and rabbit anti-P2X3 (1:300; Neuromics). Next, the membranes were washed and incubated with an alkaline phosphatase-conjugated anti-guinea pig and anti-rabbit secondary antibodies (1:200; Santa Cruz), respectively. The membranes were also incubated with mouse anti-β-actin to show equal loading of the protein. The immunoreactive proteins were detected by enhanced chemiluminescence (ECL kit). The bands recognized by the primary antibody were visualized by exposure of the membrane onto an x-ray film. Then, the film was scanned and the optical density of the bands was analyzed using the Scion Image software (National Institute of Health, USA)....

Tanja Bele and Elsa Fabbretti. The scaffold protein CASK controls ATP release in sensory ganglia upon P2X3 receptor activation and is part of an ATP keeper complex. Journal of Neurochemistry. DOI: 10.1111/jnc.13680

... X-100. The following primary antibodies were used: anti-P2X3 receptors (1:200,Alomone), guinea pig anti-P2X3 receptors (1:100; Neuromics); anti-Glutamine synthase (1:500, Sigma), anti-β-Tubulin III (1:1000, Sigma), anti ...

Saman Rahbar, Wen Pan, Michael G. Jonz. Purinergic and Cholinergic Drugs Mediate Hyperventilation in Zebrafish: Evidence from a Novel Chemical Screen. PLOS One. Published: April 21, 2016http://dx.doi.org/10.1371/journal.pone.0154261

...Zebrafish larvae at 16 dpf were prepared for immunohistochemistry following previously established procedures [23,28]. Zebrafish were euthanized with 1 mg ml-1 MS-222. Whole larvae were then fixed by immersion in a phosphate-buffered solution (PBS; 137 mM NaCl, 15.2 mM Na2HPO4, 2.7 mM KCl, 1.5 mM KH2PO4 at pH 7.8, [14]) containing 4% paraformaldehyde at 4°C overnight. After rinsing, whole larvae were permeabilized for 24-48 h using 2% Triton X-100 in PBS at 4°C.

Larvae were placed in a solution of primary antibodies, containing polyclonal rabbit anti-5-HT (1:250; cat. no. S5545, Sigma-Aldrich; Antibody ID: AB_477522) and polyclonal guinea pig anti-P2X3 (1:400-1:1,000; cat. no. GP10108; Neuromics, Edina, MN, USA; Antibody ID: AB_2283325), and diluted in 0.5% Triton X-100 at 4°C for 24 h. Tissue was then rinsed in PBS and co-treated with secondary antibodies, anti-rabbit fluorescein isothiocyanate (FITC, 1:50; cat no. 111-095-003, Jackson Immuno Research Laboratories, West Grove, PA, USA) and anti-guinea pig Alexa 594 (1:400; cat. no. A11076, Invitrogen), at room temperature for 1-2 h in the dark. Larvae were rinsed and gill complexes removed and placed on glass slides for imaging.

Polyclonal antibodies for 5-HT were raised in rabbit against a creatinine sulphate complex conjugated with BSA (manufacturer specifications). This antibody has been used to characterize the serotonergic system of gill NECs in several teleost species, including zebrafish [14,23,34]. Antibodies for P2X3 receptors were cloned from rat dorsal root ganglia and recognize residues 383-397 of the carboxy terminus of the receptor. For controls, the P2X3 antibody was pre-treated with a specific blocking peptide (1:100; cat. no. P10108, Neuromics) for 48 h at 4°C before application of the antibody to the tissue for 2 h at room temperature. This procedure eliminated P2X3 immunolabelling...

Erina Ikeda, Tetsuya Goto, Kaori Gunjigake, Kayoko Kuroishi, Masae Ueda, Shinji Kataoka, Takashi Toyono, Mitsushiro Nakatomi, Yuji Seta, Chiaki Kitamura, Tatsuji Nishihara, Tatsuo Kawamoto. Expression of Vesicular Nucleotide Transporter in Rat Odontoblasts. ACTA HISTOCHEMICA ET CYTOCHEMICA. Article ID: 15022. [Advance Publication] Released February 10, 2016. http://doi.org/10.1267/ahc.15022

...First, the sections were pre-treated with 0.1% Triton X-100 for 10 min at room temperature and pre-incubated with Blocking ONe (1:20; Nacalai, Tesque Inc., Kyoto, Japan) in 0.1 M phosphate-buffered saline (PBS, pH 7.4) for 20 min at 37°C. The sections were then incubated with mouse monoclonal antibodies against nestin (1:100; Millipore, Billerica, MA, USA), rabbit polyclonal antibodies against Snap25 (1:500; Sigma-Aldrich) or rabbit polyclonal antibodies against NF200 (1:1000; Sigma-Aldrich) for 2 hr at 37°C. After rinsing with 0.1 M PBS, the sections were incubated with goat anti-mouse IgG 568 (1:400; Invitrogen, Carlsbad, CA, USA) or goat anti-rabbit IgG 488 (1:400; Invitrogen) for 2 hr at 37°C. After rinsing again with 0.1 M PBS, the sections were incubated with rabbit polyclonal antibodies against VNUT (1:100) or guinea pig polyclonal antibodies against P2X3 (1:250; Neuromics, Edina, MN) for 2 hr at 37°C, washed in PBS and incubated with goat anti-rabbit IgG 488 (1:400; Invitrogen) for 2 hr at 37°C. After rinsing again with 0.1 M PBS, the sections were incubated with DAPI (Vector Laboratories Inc., Burlingame, CA, USA) for 5 min at room temperature. Negative control sections were incubated with secondary antibody in buffer alone. Finally, the sections were washed in 0.1 M PBS and covered with coverslips....

Sam M. Hermes, Michael C. Andresen, Sue A. Aicher. Localization of TRPV1 and P2X3 in unmyelinated and myelinated vagal afferents in the rat. Journal of Chemical Neuroanatomy Available online 17 December 2015. doi:10.1016/j.jchemneu.2015.12.003

... in an antibody cocktail consisting of a polyclonal goat antibody directed against Griffonia
(Bandeiraea) simplicifolia lectin I (IB4) (1:1000; Vector Laboratories, Cat: AS-2104) along with
a polyclonal rabbit antibody directed against P2X3 (1:7500; Neuromics, Cat: RA10109) and a ...

Jianfei Guo, Xudong Fu, Xia Cui, Minhua Fan. Contributions of purinergic P2X3 receptors within the midbrain periaqueductal gray to diabetes-induced neuropathic pain. The Journal of Physiological Sciences November 2014.

...An equal volume of total and membrane samples was applied to SDS-PAGE. Membranes were incubated with the rabbit anti-P2X 3 primary antibody (1:1000, Neuromics, Edina, MN, USA) and goat anti-rabbit secondary antibody (1:200, Neuromics, Edina, MN, USA).

Paulino Barragán-Iglesias, Hector I. Rocha-González, Jorge Baruch Pineda-Farias, Janet Murbartián, Beatriz Godínez-Chaparro, Peter S. Reinach, Thiago M. Cunha, Fernando Q. Cunha, Vinicio Granados-Soto, Inhibition of peripheral anion exchanger 3 decreases formalin-induced pain, European Journal of Pharmacology, Available online 27 May 2014, ISSN 0014-2999, http://dx.doi.org/10.1016/j.ejphar.2014.05.029. (http://www.sciencedirect.com/science/article/pii/S0014299914003914).

...substance P (guinea pig; 1:200; Cat # GP14110; Neuromics, Edina, MN), and purinergic P2×3 receptor (guinea pig: 1:1000; Cat # GP10108 ...

Longchang Fan, Xiaowei Guan, Wei Wang, Jian-Yuan Zhao, Hongkang Zhang, Vinod Tiwari, Paul N Hoffman, Min Li and Yuan-Xiang Tao. Impaired neuropathic pain and preserved acute pain in rats overexpressing voltage-gated potassium channel subunit Kv1.2 in primary afferent neurons. Molecular Pain 2014, 10:8 doi:10.1186/1744-8069-10-8.

... rabbit anti-P2X3 (Neuromics, Edina, MN)...

Suzy Krimon, Dionéia Araldi, Filipe César do Prado, Cláudia Herrera Tambeli, Maria Cláudia G. Oliveira-Fusaro, Carlos Amílcar Parada. P2X3 receptors induced inflammatory nociception modulated by TRPA1, 5-HT3 and 5-HT1A receptors. Pharmacology Biochemistry and Behavior, Available online 8 October 2013. http://dx.doi.org/10.1016/j.pbb.2013.09.017.

...containing 5% non-fat dry milk at room temperature, followed by incubation with P2X3 rabbit polyclonal antibody (1:500; Neuromics) overnight at 4 °C, rinsed six times with TBST, and then incubated for 40 minutes in goat anti-rabbit IgG peroxidase...

Swathi K. Hullugundi,Michel D. Ferrari, Arn M. J. M. van den Maagdenberg, Andrea Nistri. Andrea Nistri. The Mechanism of Functional Up-Regulation of P2X3 Receptors of Trigeminal Sensory Neurons in a Genetic Mouse Model of Familial Hemiplegic Migraine Type 1 (FHM-1). PLoS ONE 8(4): e60677. doi:10.1371/journal.pone.0060677

...P2X3 (1:300; Neuromics, Edina, MN, USA)... Andrea Nistri

Liron S. Durakua, Mehdi Hossaini, Barthold N. Schüttenhelm, b, Joan C. Holstege, Martijn Baas, Tom J.H. Ruigrok, Erik T. Walbeehm. Re-innervation patterns by peptidergic Substance-P, non-peptidergic P2X3, and myelinated NF-200 nerve fibers in epidermis and dermis of rats with neuropathic pain. Experimental Neurology Volume 241, March 2013, Pages 13–24. doi.org/10.1016/j.expneurol.2012.11.029.

... P2X3 (1/25.000, guinea pig, Neuromics)...

Gary Mo, Jennifer C. Peleshok, Chang-Qing Cao, Alfredo Ribeiro-da-Silva and Philippe Séguéla. Control of P2X3 channel function by metabotropic P2Y2 UTP receptors in primary sensory neurons. Molecular Pharmacology Fast Forward. Published on December 18, 2012 as doi:10.1124/mol.112.082099.

Dissociated DRG neurons were cultured on glass coverslips for 24 h prior to fixation with 4% PFA. Slides were washed for 30 min with PBS followed by 1 h incubation with 10% blocking buffer. Slides were then incubated overnight at 4,sup>oC in humidified chamber with the following primary polyclonal antibodies: guinea-pig anti-P2X3 (1:7000, Neuromics, Edina MN)

F.C. Pradoa, D. Araldia, A.S. Vieiraa, M.C.G. Oliveira-Fusarob, C.H. Tambelia, C.A. Parada. Neuronal P2X3 receptor activation is essential to the hyperalgesia induced by prostaglandins and sympathomimetic amines released during inflammation. http://dx.doi.org/10.1016/j.neuropharm.2012.11.011.

......non-fat dry milk at room temperature, followed by incubation with P2X3 or PKCɛ rabbit polyclonal antibody (1:500; Neuromics) overnight at 4 °C, rinsed six times with TBST, and then incubated for 40 min in goat anti-rabbit IgG peroxidase conjugate...

Min Liu, PhD, MD, Yun-fei Xu, PhD, MD, Yuan Feng, PhD, MD, Fengqiang Yang, MD, Jun Luo, MD, Wei Zhai, PhD, MD, Jian-ping Che, MD, Guang-chun Wang, MD, Jun-hua Zheng, PhD. Epigallocatechin gallate attenuates interstitial cystitis in human bladder urothelium cells by modulating purinergic receptors. Journal of Surgical Research. http://dx.doi.org/10.1016/j.jss.2012.11.041.

...P2X2 (Neuromics, Northfield, MN)...

Abeer W Saeed, Alfredo Ribeiro-da-Silva. Non-peptidergic primary afferents are presynaptic to neurokinin-1 receptor immunoreactive lamina I projection neurons in rat spinal cord. Molecular Pain 2012, 8:64 doi:10.1186/1744-8069-8-64.

...rabbit anti-CGRP and guinea pig anti-P2X3 at a 1:25,000 dilution (Neuromics, Edina, MN)...

Anna M.W. Taylora, Maria Osikowicza, Alfredo Ribeiro-da-Silva. Consequences of the ablation of nonpeptidergic afferents in an animal model of trigeminal neuropathic pain. PAIN. Volume 153, Issue 6, June 2012, Pages 1311–1319. http://dx.doi.org/10.1016/j.pain.2012.03.023.

...Sections were then incubated for 48 hours at 4°C with a guinea pig polyclonal anti-P2X 3 antibody (1:25,000; Neuromics, Edina, MN, USA), diluted in PBS-T. Following primary antibody incubation, sections were treated with a biotin-conjugated...

T. Cho, V. V. Chaban. Interaction Between P2X3 and Oestrogen Receptor (ER)α/ERβ in ATP-Mediated Calcium Signalling In Mice Sensory Neurones. Journal of Neuroendocrinology Volume 24, Issue 5, pages 789–797, May 2012

...with polyclonal rabbit antibody against P2X3 receptor (dilution 1 : 1000; Neuromics)...

Ji Z-G , Ito S , Honjoh T , Ohta H , Ishizuka T , et al. 2012 Light-evoked Somatosensory Perception of Transgenic Rats That Express Channelrhodopsin-2 in Dorsal Root Ganglion Cells. PLoS ONE 7(3): e32699.

 ...guinea-pig anti-P2X3 (1:1,000, GP10108, Neuromics, Edina, MN, USA)...

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)...

B. Huanga, X. Zhaoc, L.-B. Zhengb, L. Zhanga, B. Nia. Different expression of tissue inhibitor of metalloproteinase family members in rat dorsal root ganglia and their changes after peripheral nerve injury. Neuroscience, Volume 193, 13 October 2011, Pages 421-428.

...anti-P2X3 (rabbit, Neuromics, MN, USA)...

Gabriela Castañeda-Corral, Héctor I. Rocha-González, Beatriz Godínez-Chaparro, Juan Miguel Jiménez-Andrade and Vinicio Granados-Soto. Role of the spinal Na+/H+ exchanger in formalin-induced nociception. Neuroscience Letters. doi:10.1016/j.neulet.2011.06.048.

...SP (guinea pig; 1:500; Cat# GP14110; Neuromics), CGRP (goat, 1:500; Cat# Ab36001; Abcam) and P2X3 receptor (guinea pig: 1:10,000; Cat# GP10108; Neuromics)...

Anna M.W. Taylora and Alfredo Ribeiro-da-Silva. GDNF levels in the lower lip skin in a rat model of trigeminal neuropathic pain: Implications for nonpeptidergic fiber reinnervation and parasympathetic sprouting. PAIN Volume 152, Issue 7, July 2011, Pages 1502-1510. doi:10.1016/j.pain.2011.02.035.

...Sections were then incubated for 48h at 4°C with a guinea pig polyclonal anti-P2X3 (1:25,000; Neuromics, Edina, MN)...

Jennifer C. Peleshok, Alfredo Ribeiro-da-Silva. Delayed reinnervation by nonpeptidergic nociceptive afferents of the glabrous skin of the rat hindpaw in a neuropathic pain model. The Journal of Comparative Neurology. Volume 519, Issue 1, pages 49–63, 1 January 2011.

...The polyclonal anti-P2X3 antibody was raised in guinea pig (Neuromics, GP10108)...

C.K. Park, J.H. Bae, H.Y. Kim, H.J. Jo, Y.H. Kim, S.J. Jung, J.S. Kim, and S.B. Oh. Substance P Sensitizes P2X3 in Nociceptive Trigeminal Neurons. Journal of Dental Research, October 2010; 89: 1154 - 1159.

...The sections were incubated overnight at 4C with a mixture of guinea pig anti-P2X3 antibody (1:500; Neuromics, Minneapolis, MN, USA) and rabbit anti-NK-1 antibody (1:1000; Chemicon, Temecula, CA, USA). The sections were then incubated...

Timothy K. Y. Kaan, Ping K. Yip, Sital Patel, Meirion Davies, Fabien Marchand, Debra A. Cockayne, Philip A. Nunn, Anthony H. Dickenson, Anthony P. D. W. Ford, Yu Zhong, Marzia Malcangio, and Stephen B. McMahon Systemic blockade of P2X3 and P2X2/3 receptors attenuates bone cancer pain behaviour in rats. Brain, September 2010; 133: 2549 - 2564.

......Slides were then incubated with rabbit anti-P2X3 (1:2000, Neuromics) and sheep anti-calcitonin gene-related peptide (1:1000, Biomol...anti-beta-III-tubulin (1:4000, Promega) and guinea pig anti-P2X3 (1:100, Neuromics). The next day, after three washes with phosphate-buffered......

James P. Lund,Somayeh Sadeghi,Tuija Athanassiadis, Nadia Caram Salas, François Auclair, Benoît Thivierge, Isabel Arsenault, Pierre Rompré, Karl-Gunnar Westberg, and Arlette Kolta.Assessment of the Potential Role of Muscle Spindle Mechanoreceptor Afferents in Chronic Muscle Pain in the Rat Masseter Muscle. PLoS One. 2010; 5(6): e11131. Published online 2010 June 15. doi: 10.1371/journal.pone.0011131.

Reagents Referenced:

Nathaniel A. Sowa, Bonnie Taylor-Blake, and Mark J. Zylka. Ecto-5'-Nucleotidase (CD73) Inhibits Nociception by Hydrolyzing AMP to Adenosine in Nociceptive Circuits. The Journal of Neuroscience, February 10, 2010, 30(6):2235-2244; doi:10.1523/JNEUROSCI.5324-09.2010. 

...rabbit anti-P2X3-RA10109, Neuromics; 1:750), rabbit anti- VR1 C-Terminus (TRPV1) - mouse specific (RA14113, Neuromics; 1:750)...

Xu GY, Winston JH, Shenoy M, Zhou S, Chen JD Z, Pasricha PJ. The endogenous hydrogen sulfide producing enzyme cystathionine-beta synthase contributes to visceral hypersensitivity in a rat model of irritable bowel syndrome. Molecular Pain 2009, 5:44 (6 August 2009).

...For triple labeling, sections were simultaneously incubated with TRPV1 (1:200, Santa Cruz, CA) [19] or P2X3R (1:200, Neuromics, MN) [17] antibody and then incubated with Alexa Fluor 355. Negative control was performed by omitting the primary antibody. Sections were viewed with filter cubes appropriate for DiI (rhodamine filter), Alexa 488 and 355. Images were captured and analyzed using Metaview software (Nikon, Melville, NY). To ensure that a neuron was counted only once, serial sections were placed on consecutive slides with at least 50 μm between sections on the same slide... 

Michael A. Thacker, Anna K. Clark, Thomas Bishop, John Grist , Ping K. Yip, Lawrence D.F. Moon, Stephen W.N. Thompson , Fabien Marchand , Stephen B. McMahon. CCL2 is a key mediator of microglia activation in neuropathic pain states. European Journal of Pain 13 (2009) 263–272

...P2X3(1:500, rabbit anti-P2X3, marker for small purinergic fibres; Neuromics, USA)...

Marianna D'Arco, Rashid Giniatullin, Vanessa Leone, Paolo Carloni, Nicol Birsa, Asha Nair, Andrea Nistri, and Elsa Fabbretti The C-terminal Src Inhibitory Kinase (Csk)-mediated Tyrosine Phosphorylation Is a Novel Molecular Mechanism to Limit P2X3 Receptor Function in Mouse Sensory Neurons J. Biol. Chem., Aug 2009; 284: 21393 - 21401.

...Paraformaldehyde fixed trigeminal neurons were processed with a guinea pig anti-P2X3 antibody (1:400, Neuromics, Edina, MN)....

Anna M.W. Taylor, Jennifer C. Peleshok, Alfredo Ribeiro-da-Silva. Distribution of P2X3-immunoreactive fibers in hairy and glabrous skin of the rat. Comp. Neurol. 514:555-566, 2009. © 2009 Wiley-Liss, Inc.

...P2X3 antibody (Neuromics; GP10108, lot 400457)...

Elsa Fabbretti*, Elena Sokolova*, Lara Masten, Marianna D’Arco, Alessandra Fabbro, Andrea Nistri and Rashid Giniatullin. Identification of negative residues in the P2X3 ATP receptor ectodomain as structural determinants for desensitization and the Ca2+ sensing modulatory sites. JBC Papers in Press. Published on October 8, 2004 as Manuscript M409772200.

...Western immunoblots of transfected or untransfected HEK cells were performed as recently reported (19). In brief, these were lysed using a buffer containing 100 mM Tris HCl (pH 6.8), 200 mM dithiothreitol, 4 % SDS, 20 % glycerol and a cocktail of protease inhibitors (Sigma), and separated on 10 % polyacrylamide gel. After blocking with Tris saline buffer (TBS) containing milk, Tween 20 and preimmune serum, proteins were incubated overnight at 4°C with an anti-P2X3 antibody (Neuromics; 1:2000). Immunocomplexes were incubated for 1 h with a peroxidase-conjugated secondary antibody (Sigma; 1:4000) and detected with a chemiluminescence ECL kit (Amersham). Negative controls were obtained by mock transfection of HEK cells with the pEGFP-N1 plasmid (CLONTECH). Controls for efficient loading of different lysate materials were carried out by using an anti-β actin antibody (mouse monoclonal, 1:2000, Sigma)...

Sylvia Sarman; Jorge Mancini; Ingeborg van der Ploeg; J. Oscar Croxatto; Anders Kvanta; Juan E. Gallo. Involvement of Purinergic P2 Receptors in Experimental Retinal Neovascularization. Current Eye Research. 10.1080/02713680701885470 .

Five eyes (5 animals) from either hyperoxia- or non-hyperoxia-treated mice were enucleated and fixed for 48 hr at 4°C in paraformaldehyde (Sigma-Aldrich, St Louis, MO, USA). Afterward, eyes were immersed for cryoprotection in graded sucrose solution (5% overnight, 10%, 15%, and 20%) and interlocked with resin. Fourteen micron sections were obtained (Shandon AS325 Retraction Microtome, Thermo Scientific, Waltman, MA, USA) and fixed on polylysine-treated glass slides. After overnight incubation in primary antibody (P2X1 1:1000,  P2X2 1:1000,P2X3  1:1000, and  P2Y2 1:800) (Neuromics, Minneapolis, MN, USA) at 4°C, sections were treated with biotinylated secondary antibodies followed by an avidin peroxidase complex step (Vectastin Elite ABC, Vector Laboratories, Burlingame, CA, USA). Finally, a color reaction was obtained using 3,3'-diaminobenzidine (DAB)/nickel-enhanced solution for staining (Sigma-Aldrich, St Louis, MO, USA). At least 6 sections per retina were analyzed. For immunofluorescence, the sections were labeled with lissamine rhodamine-conjugated goat anti-rabbit Ig-G or with fluorescein-5-isothiocyanate-conjugated goat anti-guinea pig Ig-G (Jackson Immunoresearch Laboratories, West Grove, PA, USA). At least 6 sections per retina were analyzed. Visualization was done using a Nikon Fluorescence Eclipse Microscope (Tokyo, Japan), and photographs were taken with a Nikon DN 100 Digital Camera (Tokyo, Japan).

Loddenkemper, Walter Zidek, Joachim Jankowski and Markus van der Giet, Franziska Hub, Joanna Kowalska, Jacek Jemielity, Andrzej Guranowski, Christoph.  Markus Tölle, Vera Jankowski, Mirjam Schuchardt, Annette Wiedon, Tao Huang. Adenosine 5'-Tetraphosphate Is a Highly Potent Purinergic Endothelium-Derived Vasoconstrictor.  DOI: 10.1161/CIRCRESAHA.108.177865.

For immunostaining, 2-3μm thick serial sections were cut, deparaffinized and subjected to a heat-induced epitope retrieval step before incubation with antibodies. Sections were immersed in sodium citrate buffer solutions at pH 6.0 and heated in a high-pressure cooker. The slides were rinsed in cool running water, washed in Tris-buffered saline (pH 7.4), and incubated with primary antibodies. The primary antibodies included rabbit polyclonal antibodies against P2X1 (Neuromics, Edina, Minnesota, USA, dilution 1:5000).

Ruizhong Wang, Tamara King, Michael H Ossipov, Anthony J Rossomando, Todd W Vanderah, Pamela Harvey, Peter Cariani, Eric Frank, Dinah W Y Sah, Frank Porreca. Persistent restoration of sensory function by immediate or delayed systemic artemin after dorsal root injury.  Published online: 23 March 2008 | doi :10.1038/nn2069

...P2X3 (host rabbit/guinea pig, 1:10,000, Neuromics)...

Guang-Yin Xu, Mohan Shenoy, John H Winston, Sahil Mittal  and P Jay Pasricha. P2X receptor-mediated visceral hyperalgesia in a rat model of chronic visceral hypersensitivity. Gut. Published Online First: 12 February 2008. doi:10.1136/gut.2007.134221.

...then incubated overnight at 4°C with antibodies against P2X 3 (Neuromics...

Elsa Fabbretti, Marianna D'Arco, Alessandra Fabbro, Manuela Simonetti, Andrea Nistri, and Rashid Giniatullin. Delayed Upregulation of ATP P2X3 Receptors of Trigeminal Sensory Neurons by Calcitonin Gene-Related Peptide The Journal of Neuroscience, June 7, 2006, 26(23):6163-6171

... immunoblot using antibodies against the P2X3 receptor (dilution 1:2000; Neuromics, Edina, ...

Min Liu, Wenlong Huang, Dongsheng Wu and John V. Priestley. TRPV1, but not P2X3, requires cholesterol for its function and membrane expression in rat nociceptors . European Journal of Neuroscience Volume 24 Page 1 - July 2006

...guinea-pig P2X3 antibody (1 : 5000, Neuromics) overnight...

Carole Torsney, Rebecca L Anderson, Kerry-Anne G Ryce-Paul, and Amy B MacDermott. Characterization of sensory neuron subpopulations selectively expressing green fluorescent protein in phosphodiesterase 1C BAC transgenic mice Mol Pain. 2006; 2: 17.  Published online 2006 May 8. doi: 10.1186/1744-8069-2-17

...guinea-pig antiserum to P2X3 receptor (1:20000, Neuromics, ...

Torsney C, Anderson RL, Ryce-Paul KAG, MacDermott AB. Characterization of sensory neuron subpopulations selectively expressing green fluorescent protein in phosphodiesterase 1C BAC transgenic mice Molecular Pain 2006, 2:17 (8 May 2006)

...IHC: guinea-pig antiserum to P2X3 receptor (1:20,000, Neuromics, Northfield, MN)...

Dorn G, Patel S, Wotherspoon G, Hemmings-Mieszczak M, Barclay J, Natt FJ, Martin P, Bevan S, Fox A, Ganju P, Wishart W, Hall J. siRNA relieves chronic neuropathic pain.  Nucleic Acids Res. 2004 Mar 16;32(5):e49. 

Honore P, Kage K, Mikusa J, Watt AT, Johnston JF, Wyatt JR, Faltynek CR, Jarvis MF, Lynch K. Analgesic profile of intrathecal P2X(3) antisense oligonucleotide treatment in chronic inflammatory and neuropathic pain states in rats. Pain. 2002 Sep;99(1-2):11-9. 

Sun, Y, Chai, T. Up-Regulation of P2X3 Receptor During Stretch of Bladder Urothelial Cells From Patients With Interstitial Cystitis.

Journal of Urology. 171(1):448-452, January 2004 

Hubscher, Charles H.; Petruska, Jeffrey C.; Rau, Kristofer K.; Johnson, Richard D. Co-expression of P2X receptor subunits on rat nodose neurons that bind the isolectin GS-I-B4

Neuroreport. 12(13):2995-2997, September 17, 2001.

Fabbretti E, Sokolova E, Masten L, D'Arco M, Fabbro A, Nistri A, and Giniatullin R. Identification of Negative Residues in the P2X3 ATP Receptor Ectodomain as Structural Determinants for Desensitization and the Ca2+-sensing Modulatory Sites.  J. Biol. Chem., Vol. 279, Issue 51, 53109-53115

...the antibody against P2X3 was from Neuromics...

Averill S, Michael G, Shortland P, Leavesley R, King V, Bradbury E, McMahon S, and Priestley J. NGF and GDNF ameliorate the increase in ATF3 expression which occurs in dorsal root ganglion cells in response to peripheral nerve injury.  Eur. J. Neurosci. 19(6):1437-1445

...guinea pig anti-P2X3 (Neuromics... 

Sokolova E, Skorinkin A, Fabbretti E, Masten L, Nistri A, and Giniatullin R. (2004) Agonist-dependence of recovery from desensitization of P2X3 receptors provides a novel and sensitive approach for their rapid up or downregulation.  British J. of Pharmaco. (2004) 141, 1048–1058

...For Western immunoblots... and then incubated overnight at 41C with antibodies against P2X3 (Neuromics; 1:1000)...

Guang-Yin Xu and Li-Yen Mae Huang. (2004). Ca2++/calmodulin-dependent protein kinase II.PNAS, 101(32): 11868–11873

...anti-P2X3 receptors (1:1,000; Neuromics... 

Fu X, Nurse C, and Cutz E. (2004). Expression of functional purinergic receptors in pulmonary neuroepithelial bodies and their role in hypoxia chemotransmission.  Biol. Chem. 385(3-4), p275. 

Averill S, Davis DR*, Shortland PJ, Priestley JV, Hunt SP. Dynamic Pattern of Reg-2 Expression in Rat Sensory Neurons after Peripheral Nerve Injury. J. Neurosci., 22(17):7493-7501 

Nagaya N, Tittle RK, Saar N, Dellal SS, Hume RI. AN INTERSUBUNIT ZINC BINDING SITE IN RAT P2X2 RECEPTORS.  J Biol Chem. 2005 July 15; 280(28): 25982–25993.

...Blots were then probed a rabbit polyclonal antibody directed against residues 460–472 of the rat P2X2 receptor (Neuromics, Bloomington, MN) and visualized by ECL (Amersham, Piscataway, NJ).

Hiroyuki Ichikawa, and Tomosada Sugimoto. The co-expression of P2X3 receptor with VR1 and VRL-1 in the rat trigeminal ganglion.  Brain Research Volume 998, Issue 1, 13 February 2004, Pages 130-135

...VR1- and VRL-1-containing neurons co-express CGRP, supply the skin with free ... receptor sera were preabsorbed with the protein (20 μg/ml, Neuromics)...