|Calculated MW||60074 Da|
|Homology||Mouse - 16/18 amino acid residues identical; rat - 14/18 amino acid residues identical.|
|Other Names||Muscarinic acetylcholine receptor M5, CHRM5|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)HNATTVNGTPVNHQPLER, corresponding to amino acid residues 7-24 of human M5 muscarinic receptor (Accession P08912). Extracellular, N-terminus.|
|Peptide Confirmation||Confirmed by amino acid analysis.|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 mg/ml.|
|Buffer After Reconstitution||Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.|
|Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).|
|Control Antigen Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Control Antigen Reconstitution||100 µl water.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||1 µg peptide per 1 µg antibody.|
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Provided below are standard protocols that you may find useful for product applications.
Acetylcholine is a neurotransmitter which activates two different groups of receptors: nicotinic acetylcholine receptors (nAChRs) which belong to the superfamily of ligand-gated ion channels and muscarinic acetylcholine receptors (mAChR), belonging to the G-protein coupled receptor (GPCR) superfamily. There are five separate gene products that put together this GPCR subfamily, M1-M5, and like all GPCR have seven transmembrane spanning domains. Generally, M1, M3 and M5 muscarinic receptors are known to activate phospholipase C (PLC) via Gq coupling, while M2 and M4 muscarinic receptors couple to Gi/o, and therefore inhibit adenylate cyclase1. This classification is however not clear cut. mAChR can activate adenylate cyclase by coupling to Gs2. In addition, M2 and M4 muscarinic receptors, when overexpressed can also activate adenylate cyclase in some systems3-5. Evidence suggests that muscarinic receptors can form homo or heterodimers at that the dimer formed can subsequently affect the downstream signaling pathways1. Muscarinic receptors have been shown to regulate voltage-gated Ca2+ channels, namely Cav2.1 6 as well acid sensing ion channels (ASIC)7. Generally speaking the actions of muscarinic receptors on ion channels can be either via 2nd messengers or through their direct action on the channels once activated8. Furthermore, muscarinic receptors can also promote endocytosis of ion channels (Kv1.2 for example) by recruiting tyrosine kinases that phosphorylate the channel in order to terminate its activity9. Other diverse and important functions of muscarinic receptors include cell growth, survival and physiology1. Expression of muscarinic receptors is found in neurons in the central nervous system as well as in the peripheral nervous system. In the non-nervous system, these receptors are expressed in the cardiac and smooth muscle, lung, intestine, ovary and urothelium1,10. M5 muscarinic receptor has been the hardest receptor to study as there is no specific pharmacology targeted against the receptor and no high level of expression has been detected in tissues. However, the effects of single knockout mice were studied and it was observed that in M5 muscarinic receptor knockout mice, there is a moderate decrease in ligand binding in the lung and bladder, two organs expressing M5 muscarinic receptor11. In addition, it has been proposed that M5 muscarinic receptor may be involved in interstitial cystitis, a urinary bladder disorder12. Abgent is pleased to offer a highly specific antibody directed against an extracellular epitope of human M5 muscarinic receptor. Anti-M5 Muscarinic Receptor (extracellular) antibody (#AG1423) can be used for western blot analysis. It was specifically designed to recognize rat, mouse and human M5 muscarinic receptor.
1. Nathanson, N.M. (2008) Pharmacol. Ther. 119, 33.
2. Migeon, J.C. and Nathanson N.M. (1994) J. Biol. Chem. 269, 9767.
3. Ashkenazi, A. et al. (1987) Science 238, 672.
4. Tietje, K.M. et al. (1990) J. Biol. Chem. 265, 2828.
5. Katz, A. et al. (1992) Nature 360, 686.
6. Perez-Burgos, A. et al. (2010) Neuroscience 165, 293.
7. Dorofeeva, N.A. et al. (2009) Neuroreport 20, 1386.
8. Wickman, K. and Clapham, D.E. (1995) Physiol. Rev. 75, 865.
9. Nesti, E. et al. (2004) Mol. Biol. Cell 15, 4073.
10. Wessler, I. and Kirkpatrick, C.J. (2008) Br. J. Pharmacol. 154, 1558.
11. Ito, Y. et al. (2009) Br. J. Pharmacol. 156, 1147.
12. Giglio, D. et al. (2005) Auton. Neurosci. 122, 9.
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