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5-HT (Serotonin) 2A Receptor Antibody

24288
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  • Refer to Dr. Schwendt's review
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Price: $375.00
Product ID : 24288
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Description

The antibody is provided as 100 uL of affinity purified serum in PBS (0.02 M sodium phosphate with 0.15 M sodium chloride, pH 7.5) with 1% BSA (bovine serum albumin), and 0.02% sodium azide.  Reacts with mouse, rat, and rabbit species.

The images above labeled "ImmunoStar Rabbit 5-HT 2A receptor antibody" are the results of staining of the mouse 5-HT 2A receptor in the brain, courtesy of Dr. Magdalena Zaniewska, Max-Delbruck-Centrum fur Molekulare Medizin Berlin, Germany.

The ImmunoStar 5HT 2A receptor antibody was quality control tested using standard immunohistochemical methods. The antiserum demonstrates strongly positive labeling of rat cortex, amygdala and hippocampus using indirect immunofluorescent and biotin/avidin-HRP techniques. Recommended primary dilutions are 1/300 - 1/500 in PBS/0.3% Triton X-100 - Bn/Av-HRP Technique . The addition of intensifying reagents such as nickel ammonium sulfate to the chromogen solution will approximately double the dilution factor as recommended.

Immunolabeling is completely abolished by preadsorption with synthetic rat 5HT2A receptor (22-41). Immunolabeling of Western blot revealed a single band of approximately 53kD. Due to the difficulty in running westerns with this antibody, ImmunoStar does not support or warrant the western blot application.  The western information is included as specificity information only.

Photo Description:  Low magnification IHC image of neurons staining for the 5-HT2A receptor in the rat cortex (top of page) and image of neuronal expression of the receptor in the amygdala (below). The bottom right photo is of the cortex.  The tissue was fixed with 4% formaldehyde in 0.1 M phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:500 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen.

5HT2A Amygdala #24288   5HT2A Cortex, #24288
   

Host: Rabbit

Quantity / Volume: 100 uL

State: Liquid

Reacts With: Human, Marine, Mouse, Rabbit, Rat

Alternate Names:  5-hydroxytryptamine receptor 2A; 5Ht-2; 5-hydroxytryptamine (serotonin) receptor 2A, G protein-coupled

Gene Symbol / ID, Accession #:  Htr2a,29595

 

Download Product Spec Sheet: 5-HT (Serotonin) 2A Receptor Antibody

 

Journal References:


These publications reference ImmunoStar's antibody product line, formerly sold by Diasorin, Incstar, and ImmunoNuclear.

Fay, Richard, and Leszek Kubin. "5-HT 2A receptor-like protein is present in small neurons located in rat mesopontine cholinergic nuclei, but absent from cholinergic neurons." Neuroscience letters 314.1 (2001): 77-81. Tested in Rat, PubMed ID: 11698151

Visual Neurosci,16:221-230, 5-HT2a receptors in the rabbit retina: Potential presynaptic modulators. (1999) Pootanakit K, Prior KJ, Hunter DD, Brunken WJ, IHC method, Tested in Rabbit, PubMed ID: 10367957

Philippe Coulon,Tatyana Kanyshkova,Tilman Broicher, Thomas Munsch, Nina  Wettschureck, Thomas Seidenbecher, Sven G. Meuth, Stefan Offermanns, Hans-Christian Pape and Thomas Budde, Activity modes in thalamocortical relay neurons are modulated by Gq/G11 family G-proteins – serotonergic and glutamatergic signaling, Cellular Neuroscience,4:132, Tested in Mouse, PubMed ID: 21267426

Kwan, Marilyn L., et al. "Airway vasculature after mycoplasma infection: chronic leakiness and selective hypersensitivity to substance P." American Journal of Physiology-Lung Cellular and Molecular Physiology 280.2 (2001): L286-L297.  Tested in Rat, PubMed ID: 11159008

Vázquez, Delia M., et al. "Brain 5-HT receptor system in the stressed infant rat: implications for vulnerability to substance abuse." Psychoneuroendocrinology 27.1 (2002): 245-272. Vázquez DM, Eskandari R, Zimmer CA, Levine S, López JF.  Tested in Rat, PubMed ID: 11750782    

Itoh, Takeo, and Junko Kajikuri. "Characteristics of the actions by which 5‐HT affects electrical and mechanical activities in rabbit jugular vein." British journal of pharmacology 164.3 (2011): 979-991. Takeo Itoh, Junko Kajikuri, Tested in Rabbit, PubMed ID: 21449974

Maekawa, Takashi, et al. "Characteristics of the actions by which 5‐hydroxytryptamine affects electrical and mechanical activities in rabbit jugular vein graft." British journal of pharmacology 166.4 (2012): 1419-1432. Maekawa T, Komori K, Kajikuri J, Itoh T. Tested in Rabbit, IF method, PubMed ID: 22251164

Rasul, Aram, et al. "Chronic mild stress modulates 5-HT1A and 5-HT2A receptor expression in the cerebellar cortex of NC/Nga atopic-like mice." Archives of dermatological research 305.5 (2013): 407-413.    Rasul A, Johansson B, Lonne-Rahm SB, Nordlind K, Theodorsson E, El-Nour H. Tested in Mouse, IHC method, PubMed ID: 23440396

Journal Of Vascular Surgery, 50, 3:617-625 Chronic treatment of hydroxytryptamine type 2a receptor (2009), Akio Kodama, MD, PhD, Kimihiro Komori, MD, PhD, Junko Kajikuri, PhD, and Takeo Itoh, PhD. Tested in Rabbit, IF method, PubMed ID: 19700096

Volgin, Denys V., Georg M. Stettner, and Leszek Kubin. "Circadian dependence of receptors that mediate wake-related excitatory drive to hypoglossal motoneurons." Respiratory physiology & neurobiology 188.3 (2013): 301-307.    Circadian dependence of receptors that mediate wake-related excitatory drive to hypoglossal motoneurons, PubMed ID: 23665050

Pearlstein, E., et al. "Contribution of 5‐HT to locomotion–the paradox of Pet‐1−/− mice." European Journal of Neuroscience 33.10 (2011): 1812-1822. Pearlstein E, Bras H, Deneris ES, Vinay L. Tested in Mouse, PubMed ID: 21501257

Zeng, Fanqing, Chen Xu, and Ge Xu. "Distribution of serotonin 5-HT2A and 5-HT7 receptors in the Onuf's nucleus of the rat spinal cord." Neural Regen Res 3.8 (2008): 853-857. Tested in Rat.

Rasul, Aram, et al. "Effect of chronic mild stress on serotonergic markers in the skin and brain of the NC/Nga atopic-like mouse strain." Archives of dermatological research 303.9 (2011): 625-633. Rasul A, El-Nour H, Blakely RD, Lonne-Rahm SB, Forsberg J, Johansson B,Theodorsson E, Nordlind K. Tested in Mouse, PubMed ID: 21400247

Hoshi University,1412:RP. 1-6 (2009) Enhancement of mesenteric artery contraction to 5-HT depends on Rho kinase     Takayuki Matsumoto, TsuneoKobayashi, Keiko Ishida, Kumiko Taguchi and Katsuo Kamata, Tested in Mouse, WB method, PubMed ID: 20590603

Sławińska, Urszula, et al. "Grafting of fetal brainstem 5-HT neurons into the sublesional spinal cord of paraplegic rats restores coordinated hindlimb locomotion." Experimental neurology 247 (2013): 572-581. Sławińska U, Miazga K, Cabaj AM, Leszczyńska AN, Majczyński H, Nagy JI, JordanLM. Tested in Rat, PubMed ID: 23481546

Weber, Elaine T., and Rodrigo Andrade. "Htr2a gene and 5-HT2A receptor expression in the cerebral cortex studied using genetically modified mice." Frontiers in neuroscience 4 (2010). Weber ET, Andrade R., Tested in Mouse, PubMed ID: 20802802

Hayashi, Takayuki, et al. "Intracerebroventricular fluvoxamine administration inhibited pain behavior but increased Fos expression in affective pain pathways." Pharmacology Biochemistry and Behavior 91.3 (2009): 441-446. Hayashi T, Miyata M, Nagata T, Izawa Y, Kawakami Y. IHC method, PubMed ID: 18817806

Stettner, Georg M., Leszek Kubin, and Denys V. Volgin. "Loss of motoneurons in the ventral compartment of the rat hypoglossal nucleus following early postnatal exposure to alcohol." Journal of chemical neuroanatomy 52 (2013): 87-94. Stettner GM, Kubin L, Volgin DV. Tested in Rat, PubMed ID: 23932955

Fay, Richard, and Leszek Kubin. "Pontomedullary distribution of 5‐HT2A receptor‐like protein in the rat." Journal of Comparative Neurology 418.3 (2000): 323-345. Tested in Rat, PubMed ID: 10701830

Karaki, Samah, et al. "Quantitative Phosphoproteomics Unravels Biased Phosphorylation of Serotonin 5-HT2A Receptor at Ser280 by Hallucinogenic versus non-Hallucinogenic Agonists." Molecular & Cellular Proteomics (2014): mcp-M113. Karaki S, Becamel C, Murat S, Mannoury la Cour C, Millan MJ, Prezeau L, Bockaert J, Marin P, Vandermoere F.

Kong, Xiang-Yu, et al. "Robust upregulation of serotonin 2A receptors after chronic spinal transection of rats: An immunohistochemical study." Brain research 1320 (2010): 60-68. Kong XY, Wienecke J, Hultborn H, Zhang M.Tested in Rat, PubMed ID: 20085755

Van Steenwinckel, Juliette, et al. "Role of spinal serotonin 5-HT2A receptor in 2′, 3′-dideoxycytidine-induced neuropathic pain in the rat and the mouse." Pain 137.1 (2008): 66-80. Van Steenwinckel J, Brisorgueil MJ, Fischer J, Vergé D, Gingrich JA, Bourgoin S, Hamon M, Bernard R, Conrath M. Tested in Rat and Mouse, PubMed ID: 17888573

Rasul, Aram. "Serotonergic mechanisms in atopic dermatitis." (2012).

Xu, Chen, et al. "Serotonin 5‐HT2A and 5‐HT5A receptors are expressed by different motoneuron populations in rat Onuf's nucleus." Journal of Comparative Neurology 502.4 (2007): 620-634. Xu C, Giuliano F, Sun XQ, Brisorgueil MJ, Leclerc P, Vergé D, Conrath M. Tested in Rat, PubMed ID: 17394137

Thibault, Karine, et al. "Serotonin 5-HT2A receptor involvement and Fos expression at the spinal level in vincristine-induced neuropathy in the rat." Pain 140.2 (2008): 305-322. Thibault K, Van Steenwinckel J, Brisorgueil MJ, Fischer J, Hamon M, Calvino B,Conrath M. Tested in Rat, PubMed ID: 18930597

Loyd, Dayna R., et al. "Sex differences in serotonin enhancement of capsaicin-evoked calcitonin gene-related peptide release from human dental pulp." Pain 153.10 (2012): 2061-2067. Loyd DR, Sun XX, Locke EE, Salas MM, Hargreaves KM. Tested in Human, PubMed ID: 22819536

Neuropsychopharmacology,34:410-423, Stress impairs 5-HT2A receptor-mediated serotonergic facilitation of GABA release in juvenile rat basolateral amygdala, Xiaolong Jiang, Guoqiang Xing, Chunhui Yang, Ajay Verma, Lei Zhang and He Li    Tested in Rat, PubMed ID: 18536707

Dumitrascu, R., et al. "Terguride ameliorates monocrotaline-induced pulmonary hypertension in rats." European Respiratory Journal 37.5 (2011): 1104-1118.    Dumitrascu R, Kulcke C, Königshoff M, Kouri F, Yang X, Morrell N, Ghofrani HA,Weissmann N, Reiter R, Seeger W, Grimminger F, Eickelberg O, Schermuly RT,Pullamsetti SS. Tested in Rat, IHC method, PubMed ID: 20947677

Van Steenwinckel, J., et al. "The 5-HT2A receptor is mainly expressed in nociceptive sensory neurons in rat lumbar dorsal root ganglia." Neuroscience 161.3 (2009): 838-846. Van Steenwinckel J, Noghero A, Thibault K, Brisorgueil MJ, Fischer J, Conrath M. Tested in Rat, IHC method, PubMed ID: 19362128

Doly, Stéphane, et al. "The 5‐HT2A receptor is widely distributed in the rat spinal cord and mainly localized at the plasma membrane of postsynaptic neurons." Journal of Comparative Neurology 472.4 (2004): 496-511. Doly S, Madeira A, Fischer J, Brisorgueil MJ, Daval G, Bernard R, Vergé D,Conrath M. Tested in Rat, ICC method, PubMed ID: 15065122

Neurochemical Research,30:825–837 (2005), The pre-nervous serotonergic system of developing sea urchin embryos and larvae: pharmacologic and immunocytochemical evidence, Gennady A. Buznikov,  Robert E. Lyudmila A. Nikitina, Vladimir V., and Jean M. Lauder, IHC method, PubMed ID: 16187217

Kong, X-Y., et al. "The time course of serotonin 2A receptor expression after spinal transection of rats: an immunohistochemical study." Neuroscience 177 (2011): 114-126. Tested in Rat, PubMed ID: 21211552


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Product Reviews

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Reviewed by user
03/11/2014 - 10:36:22 AM
Only your AB shows a specific signal
(Please refer to the above western blot image submitted by Dr. Schwendt.)

We have been interested in detecting changes of 5-HT2a in rodent brain, primarily by immunoblotting, also by immunohistochemistry. We have tested several commercial antibodies, with only your Ab showing a specific signal, as verified by brain distribution, preadsorbtion and by a lack of signal in KO animals.

We have optimized a immunoblotting protocol which works well to detect 5-HT2a in rat brain lysates. Mouse tissue is a little bit more difficult but still possible

Immunoblotting: Rat or mouse brain samples were lysed by brief sonication in a buffer containing: 1% SDS in 1xPBS, protease inhibitors (Complete Mini, Roche). Samples were mixed with a modified Leamli buffer (containing a reducing agent; 40mM DTT), incubated at 37C for 30mins and separated by SDS-PAGE. Separated proteins were then blotted onto a PVDF membrane, blocked with 5% milk/TBS/0.1% Tween80 and probed with anti-5HT2a antibody (1:500, Immunostar #24288) followed by anti-rabbit HRP- conjugated secondary antibody (1:5000, Jackson Immuno). Signal was visualized using ECL2 kit according to manufacturer's instructions (Thermo).

Image Samples: 1 (Rat prefrontal cortex, 25ug), 2 (Rat laterodorsal thalamus, 25ug), 3 (Wt mouse cortex, 25 ug), 4 (5-HT2a KO mouse cortex, 25 ug), 5 (Wt mouse cortex, 75 ug), 6 (5-HT2a KO mouse cortex, 75 ug). As expected, sample in lane 2 showed low expression, and samples in lanes 4 and 6 showed no expression of 5-HT2a receptors. Mouse tissues obtained from Dr. Gonzalez-Maeso (Mount Sinai Hospital, NY) indicates detected 5Ht2a receptor signal.


Submitted by:

Marek Schwendt, PhD
Psychology Department
University of Florida
Gainesville, Fl


Reviewed by user
11/15/2012 - 02:37:02 PM
Antibodies are specific for mouse receptor
[The two pictures above labeled "ImmunoStar's Rabbit 5-HT 2A receptor antibody" are courtesy of Dr. Magdalena Zaniewska.]

Here are the results of staining of the mouse 5-HT 2A receptors in the brain with the use of Immunostar's antibodies (Cat no: 24288). I think the antibodies are specific for the mouse receptor (no receptor labeling in the brain of 5-HT 2A receptor knockout mouse) and can be used in a Western blot. What I already found out is that 5-HT 2A receptors in the cortex seem to be glycosylated (band around 75 kDa) since incubation with PNGaseF releases the protein that is around 50 kDa.

We received the 5-HT 2A receptor knockout animals from Dr. Joelle Adrien from another research team in Paris (see below).


Reviewed by:

Dr. Magdalena Zaniewska
Molekularbiologie von Hormonen im Herz-Kreislaufssystem
Max-Delbrϋck-Centrum fϋr Molekulare Medizin
Berlin, Germany

Dr. Joelle Adrien
Directeur de Recherches a l'Inserm
Faculte de Medecine Pierre et Marie Curie - Site Pitie Salpetriere
Paris, France