Human Muscarinic acetylcholine receptor M3(CHRM3) ELISA kit

Instructions
Code CSB-EL005383HU
Size 96T,5×96T,10×96T
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Product Details

Target Name cholinergic receptor, muscarinic 3
Alternative Names AChR ELISA Kit; ACM3_HUMAN ELISA Kit; cholinergic receptor muscarinic 3 ELISA Kit; CHRM 3 ELISA Kit; CHRM3 ELISA Kit; EGBRS ELISA Kit; HM 3 ELISA Kit; HM3 ELISA Kit; m3 muscarinic acetylcholine receptor ELISA Kit; M3 muscarinic receptor ELISA Kit; muscarinic 3 ELISA Kit; Muscarinic acetylcholine receptor M3 ELISA Kit; muscarinic cholinergic m3 receptor ELISA Kit; muscarinic m3 receptor ELISA Kit
Abbreviation CHRM3
Uniprot No. P20309
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
Detection Range 23.44 pg/mL-1500 pg/mL
Sensitivity 5.86 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Others
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%        
Three samples of known concentration were tested twenty times on one plate to assess.    
Inter-assay Precision (Precision between assays): CV%<10%        
Three samples of known concentration were tested in twenty assays to assess.      
               
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human CHRM3 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.  
  Sample Serum(n=4)    
1:1 Average % 96    
Range % 91-100    
1:2 Average % 90    
Range % 86-94    
1:4 Average % 102    
Range % 98-109    
1:8 Average % 93    
Range % 89-97    
Recovery
The recovery of human CHRM3 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.  
 
Sample Type Average % Recovery Range    
Serum (n=5) 95 90-100    
EDTA plasma (n=4) 84 80-88    
               
               
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.  
 
pg/ml OD1 OD2 Average Corrected    
1500 2.371 2.184 2.278 2.139    
750 1.460 1.515 1.488 1.349    
375 0.859 0.893 0.876 0.737    
187.5 0.482 0.459 0.471 0.332    
93.75 0.328 0.307 0.318 0.179    
46.88 0.247 0.232 0.240 0.101    
23.44 0.163 0.170 0.167 0.028    
0 0.136 0.141 0.139      
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 7-14 working days

Target Data

Function The muscarinic acetylcholine receptor mediates various cellular responses, including inhibition of adenylate cyclase, breakdown of phosphoinositides and modulation of potassium channels through the action of G proteins. Primary transducing effect is Pi turnover.
Gene References into Functions
  1. M3R activation-induced GRK2 recruitment is Ggamma subtype dependent in which Gbetagamma dimers with low cell membrane-affinity Ggamma9 exhibited a two-fold higher GRK2-recruitment compared to high affinity Ggamma3 expressing cells. PMID: 29864421
  2. Regulation of molecular clock oscillations and phagocytic activity via muscarinic Ca(2+) signaling in human retinal pigment epithelial cells. PMID: 28276525
  3. These results indicate that not H1 but M3 receptor-induced activation of p38 MAPK might contribute to the maintenance of epithelial barrier function through down-regulation of TNF-alpha signalling and activation of EGFR. PMID: 28412413
  4. Morphine-induced MOP receptor endocytosis is facilitated by concurrent M3 activation.M3 and MOP assemble in receptor heterocomplexes mainly located at the plasma membrane.M3-MOP receptor pharmacological interaction is independent of heterocomplex formation.M3 and MOP receptor heteromers disrupt upon both receptor endocytosis. PMID: 28411124
  5. this study shows that anti-M3R is significantly elevated in Sjogren's syndrome plasma in comparison with rheumatoid arthritis, systemic lupus erythematosus, and healthy controls PMID: 27460476
  6. Interacting post-muscarinic receptor signaling pathways potentiate MMP1 expression and invasion of human colon cancer cells. PMID: 28008134
  7. M3R expression plays an important role in early progression and invasion of colon neoplasia but is less important once tumors have spread. PMID: 28416748
  8. This study presents the endocytic pathways of internalization for muscarinic type 3 receptor and flotillin-1/2 in salivary gland epithelial cells. knockdown of flot-1 or -2 by flotillin-specific siRNA prevented internalization and reduced the endocytic efficiency of muscarinic type 3 receptor. PMID: 27221048
  9. M3-mAChR activation leads to enhancement of hsp expression via PKC-dependent phosphorylation of HSF1, thereby stabilizing the mutant hERG-FLAG protein. Thus, M3-mAChR activators may have a therapeutic value for patients with LQT2. PMID: 27803431
  10. There were no significant associations between CHRM3 SNPs and autonomic nervous system activity in patients with schizophrenia on high-dose antipsychotics. PMID: 27923235
  11. our findings support an oncogenic role for the M3 receptor in gastric cancer PMID: 26346168
  12. To our knowledge, this is the first genetic association study that reveals the genetic contribution of CHRM3 gene in bladder cancer etiology. PMID: 28053981
  13. The CHRM3 gene plays a role in abnormal thalamo-orbital frontal cortex functional connectivity in first-episode treatment-naive patients with schizophrenia. PMID: 26959877
  14. These findings indicate that M3 mAChR may be important therapeutic target for obstructive airway diseases, as it regulates the effects of the epithelial-derived chemokines on ASM cell migration, which results in lung remodeling. PMID: 26956674
  15. Blockading CHRM3 by shRNA or treatment with darifenacin inhibited prostate cancer growth. PMID: 26071486
  16. results suggest that the autoantibodies against peptides of the second extracellular loop of M3R are not pathogenic in vivo and they are not suitable as biomarkers for pSS diagnosis PMID: 26901532
  17. Report using biosensing techniques to monitor dynamic changes of inositol lipid pools in living cells reveals a PKC-dependent PtdIns4P increase upon EGF and M3 receptor activation. PMID: 26692031
  18. M3 AChR or AMP-activated protein kinase Small Interfering RNA abrogated the ACh-elicited the attenuation of Endoplasmic reticulum stress in endothelial cells, indicating that the salutary effects of ACh were likely mediated by M3 AChR-AMPK signaling. PMID: 26066647
  19. Suggest Gbeta4gamma1 as a modulator of M3 muscarinic receptor signaling. PMID: 25916507
  20. Cigarette smoking may contribute to this imbalance by affecting the polarization and survival of Th/Tregs through the up-regulation of MR3 and MR5. PMID: 25375131
  21. ACh-induced activation of EGFR/PI3K/AKT pathway and subsequent IL-8 upregulation may be one of the important mechanisms of M3R function PMID: 25964092
  22. an incorrect organizational structure of many mutants of hM3R provides the molecular basis for why they are poorly expressed and fail to be effectively trafficked to the cell surface. PMID: 25769304
  23. M3R preserves the endothelial barrier function through a mechanism potentially maintaining PTP1B activity, keeping the adherens junction proteins (AJPs) dephosphorylation. PMID: 24393526
  24. Results uncovered that Gaq binding to GRK2 enhances the recruitment of GRK2 to M3-ACh receptors. PMID: 25316767
  25. HM3-mediated IL8 expression through PKC-NFkappaB signaling pathways PMID: 24522860
  26. Overexpression of muscarinic receptor 3 promotes metastasis in non-small-cell lung cancer. PMID: 24419413
  27. The presence of muscarinic M3 receptor (HM3) autoantibodies may be related to the underlying disease mechanism in idiopathic generalized anhidrosis patients. PMID: 24334576
  28. The M3 receptor facilitates interaction of the VE-cadherin-based adherens junctional complex and the actin-based cytoskeleton by maintaining Rac1 activity, which regulates the interaction between IQGAP1/Rac1 and IQGAP1/beta-catenin. PMID: 24697698
  29. the M3 muscarinic receptor maximizes the efficiency of PLCbeta3 signaling beyond its canonical role as a guanine nucleotide exchange factor for Galpha. PMID: 24596086
  30. M3-mediated contractions in the M2KO strain were decreased 54% by the protein kinase C (PKC) inhibitor chelerythrine. PMID: 23786223
  31. Data indicate that high M3 muscarinic acetylcholine receptor (M3 receptor) expression correlates with poor survival in non-small cell lung cancer (NSCLC) patients, indicating that M3 receptor may be a novel antineoplastic target. PMID: 23838802
  32. Our findings clearly indicate that binding of anti-M3R autoantibodies to the receptor, which was verified by immunoprecipitation, suppresses AQP5 trafficking to the membrane and contribute to impaired fluid secretion in SjS. PMID: 23382834
  33. CHMR3 at the plasma membrane exists as stable dimeric complexes, a large fraction of which interact dynamically to form tetramers without the presence of trimers, pentamers or higher-order oligomers PMID: 23521066
  34. The acetylcholine M3 receptor was slightly decreased as compared to control in overactive bladder. PMID: 22691178
  35. Results suggest a major role of M(3) muscarinic receptors mediating the human gallbladder contraction through voltage-gated Ca(2+) channels and Rho kinase. PMID: 23227858
  36. Overall, the results of the present study suggest that TNF--alpha causes (1) the release of ACh from A549 cells, (2) the decrease in cholinesterase activity, and (3) the upregulation of M3R expression facilitating the autocrine effects of ACh on M3R. PMID: 23142559
  37. Generalized epilepsies implicates susceptibility genes: CHRM3 at 1q43, VRK2 at 2p16.1, ZEB2 at 2q22.3, SCN1A at 2q24.3 and PNPO at 17q21.32. PMID: 22949513
  38. The expression of mAChR M3 is influenced by the extent of differentiation, distant metastasis and the site of cholangiocarcinoma. It also plays a key role in the proliferation and metastasis of cholangiocarcinoma. PMID: 22893470
  39. CHRM3 and M(3) were suggested to play important roles in the pathogenesis of myopia and in the arrested progression of myopia by atropine. PMID: 22899762
  40. The Asn-7.49 is important for the interaction between M(3)R and ARF1 and also for the formation of the ARF/Rho/beta gamma signaling complex, a complex that might determine the rapid activation and desensitization of PLD.(muscarinic acetylcholine receptor M3) PMID: 22178951
  41. Mutation of this region in the M3-MR altered receptor coupling to G protein. PMID: 22466417
  42. Its cytoprotective effect was decreased by a M3R antagonist. PMID: 22511543
  43. The Asn-514-Tyr point mutation produces constitutive activation of muscarinic M3-AChR by decreasing the rate of receptor deactivation, while having minimal effect on receptor activation. PMID: 22564786
  44. Data suggest that M3 muscarinic receptor and Kir6.1 colocalize to detrusor caveolae; studies include tissue from both male and female subjects. PMID: 22410194
  45. The anti-M3RP205-220 antibody was detected in most patients with Sjogren syndrome. PMID: 22510450
  46. The study reports the results of a combined computational and site mutagenesis study designed to provide new insights into the orthosteric binding site of the human M3 muscarinic acetylcholine receptor. PMID: 22500107
  47. The data provide experimental evidence of the critical role that N-glycan chains play in determining muscarinic receptor distribution, localization, as well as PMID: 21798865
  48. Autoantibodies against the second extra-cellular loop of M3R could be involved in salivary dysfunction in patients with Sjogren's syndrome. PMID: 21877253
  49. Incubation of hSMG cells with pSS IgG (1mg/ml) significantly decreased M3R expression levels at the membrane. PMID: 22137887
  50. Structural basis of M3 muscarinic receptor dimer/oligomer formation. PMID: 21685385
  51. Muscarinic acetylcholine receptor M3(1q41-q44) homozygous frameshift mutation in familial congenital bladder malformation associated with a prune-belly-like syndrome, defining an isolated gene defect underlying this sometimes devastating disease. PMID: 22077972
  52. The results of the study suggest that the differential expression of the ADRB2 and CHRM3 genes in bronchial mucosa is associated with asthma and COPD clinical subtypes. PMID: 22192964
  53. CHRM3 polymorphisms are likely associated with disease duration in IBS. CHRM3 polymorphisms could be associated with IBS. PMID: 21940308
  54. The M3R C terminus is sufficient, and a six-amino-acid polybasic sequence distal to helix 8 ((565)KKKRRK(570)) is necessary for preassembly with G(q). PMID: 21873996
  55. In contrast to other genes reported to be associated with primary SJogren syndrome, like STAT4 and IRF5, CHRM3 has not been reported to be associated with other autoimmune diseases such as systemic lupus erythematosus or rheumatoid arthritis. PMID: 21450750
  56. Complex regulation of the TRPM8 cold receptor channel: role of arachidonic acid release following M3 muscarinic receptor stimulation. PMID: 21245133
  57. Data show that the crosstalk between ORs and the M3-R suggests that the functional coupling of ORs and the M3-R is required for robust OR activation. PMID: 21224444
  58. WNK4 kinase negatively regulates the anterograde trafficking of MR through kinase-independent mechanism. PMID: 21118716
  59. Distribution of M3 receptors in human colon. PMID: 20146726
  60. skin of patients with cholinergic urticaria CHRM3 lack of expression is divided into anhidrotic and wheal occuring hypohidrotic areas PMID: 20613776
  61. The results showed that microbial products up regulate the expression of M3 receptor in nasal mucosal immune cells that further increases the production of OX40L in nasal DCs and drives the production of TNF-alpha in nasal mucosa. PMID: 19951374
  62. M3 muscarinic acetylcholine receptor subtype as being primarily responsible for these transcription factor responses after stimulation with carbachol through activation of the ERK1/2 pathway PMID: 20519144
  63. the capacity of the human M(3) muscarinic acetylcholine receptor to exist as dimeric/oligomeric complexes at the surface of cells PMID: 20489201
  64. Use muscarinic antagonists to demonstrate that muscarinic M3 receptors mediate the excitatory effects of acetylcholine on small intestinal and colonic transit in humans. PMID: 20395537
  65. The results of co-immunoprecipitation of different scaffold and kinase proteins, provide experimental evidence for the role for the third cytoplasmic loop of the human M(3) muscarinic receptor in G-protein activation and multiprotein complex formation. PMID: 20332620
  66. muscarinic M3 receptor stimulation augments cigarette smoke extract-induced IL-8 (but not IL-6) production by airway smooth muscle PMID: 19460789
  67. Data show that exon-acquisition and alternative splicing events of CHRM3 genes were occurred through the continuous integration of transposable elements following conservation. PMID: 19669628
  68. potential role of endogenous GRK6 in the regulation of M(3) mACh receptor PMID: 11856737
  69. Stimulation of phospholipase C-epsilon by the M3 muscarinic acetylcholine receptor mediated by cyclic AMP and the GTPase Rap2B PMID: 11877431
  70. We show that activation of these receptors leads to divergent growth responses: M(2) AChR activation causes an increase in DNA synthesis, whereas M(3) AChR activation causes a dramatic decrease in DNA synthesis. PMID: 12126481
  71. both G(q) and G(11) are involved in mediating the action of the M(3) receptor on cytosolic Ca(2+) in HT29 cells PMID: 12194018
  72. Single nucleotide polymorphisms not more frequent in asthma. No nonrandom transmission of short tandem repeat polymorphism haplotypes in asthma. Nonrandom transmission of haplotypes in skin test reactivity to cockroach allergens. PMID: 12642833
  73. the conserved poly-basic region in the C-terminal tail of the M3 muscarinic receptor contributes to the ability to mediate protection against apoptotic cell death PMID: 12649280
  74. the M3 receptor displays a major ARF1-dependent route of phospholipase D1 activation with an additional ARF6-dependent pathway to PLD1 or PLD2 PMID: 12799371
  75. M3 mAChR desensitization is mediated by GRK6 in human SH-SY5Y cells, and receptor desensitization of phospholipase C signaling can be monitored in 'real-time' in single, living cells. PMID: 14573754
  76. The muscarinic acetylcholine receptor M3 is a mediator of bradycardia and bronchoconstriction due to vagal innervation. PMID: 14977875
  77. M3 up-regulates "sedentary" integrins alpha2beta1 and alpha3beta1. Inhibition of migration by M3 was mediated through Ca2+-dependent guanylyl cyclase-cyclic GMP-protein kinase G signaling pathway. PMID: 15263021
  78. (HEK) 293 cells expressing recombinant Galpha(q/11)-coupled muscarinic M3 receptors showed transient coexpression of RGS proteins PMID: 15383626
  79. demonstration of the antigenicity of a novel peptide fragment of the human muscarinic acetylcholine receptor 3 in primary Sjogren's syndrome PMID: 15725576
  80. binding of beta-arrestin-1 to muscarinic M(3) receptors requires paired stimulation of two receptor components within the same receptor dimer PMID: 15769745
  81. characterization of the muscarinic receptor in the spontaneously immortalized human keratinocyte cell line HaCaT and its role in cell migration PMID: 16113538
  82. The muscarinic Ca2+ signaling pathway is not necessarily affected by depolarization and suggest that the M3 receptor itself is not sensitive to voltage. PMID: 17005862
  83. C allele as associated with a reduced acute insulin response and modestly associated with increased risk of early-onset type 2 diabetes in Pima Indians. PMID: 17130513
  84. affects endocrine and exocrine hormone and salivary secretion PMID: 17192665
  85. alphaM3 belongs to a complex and diverse set of synchronously moving parts that change structure relatively late in the channel-opening process. PMID: 17513382
  86. M3R was involved in the up-regulation of H1R by activating H1R gene transcription through a PKC-dependent process PMID: 17637176
  87. Poly I:C caused increased M3R in airway smooth muscle cells by interacting with TLR3. PMID: 17851256
  88. These results indicate caveolae and caveolin-1 facilitate [Ca(2+)](i) mobilization leading to ASM contraction induced by submaximal concentrations of acetylcholine. PMID: 17890325
  89. the M(3)/M(5) subtypes appear to be the major contributor, leading to intracellular calcium mobilization from the internal store via an IP(3)-dependent pathway in the undifferentiated retinoblastoma cells. PMID: 17951979
  90. M3 employs two distinct mechanisms of chemical imitation to potently sequester chemokines, thereby inhibiting chemokine receptor binding events as well as the formation of chemotactic gradients necessary for directed leukocyte trafficking PMID: 18070938
  91. Neither the linear M3R peptide nor M3R transfectants represent suitable tools for discrimination of pathogenic from natural autoantibodies in Sjogren's syndrome. PMID: 18249005
  92. Muscarinic receptor M3 expression is associated with invasive migration of melanomas. PMID: 18422974
  93. the beta2 adrenergic and M3 muscarinic receptors, enter cells constitutively by clathrin-independent endocytosis and colocalize with markers of this endosomal pathway on recycling tubular endosomes subsequently recycle back to the plasma membrane PMID: 19033440
  94. The DEP domain of RGS7 can directly bind to the third intracellular loop of the M3R and attenuate receptor-induced Ca2+ mobilization in a M3 subtype-selective manner. PMID: 19182865
  95. Data show that NALCN and M3R belong to the same protein complex, involving the intracellular I-II loop of NALCN and the intracellular i3 loop of M3R. PMID: 19575010
  96. M(3)-muscarinic receptor can regulate the apoptotic properties of a chemotherapeutic DNA-damaging agent by regulating the expression, subcellular trafficking and modification of p53 in a manner that is, in part, dependent on the cell type. PMID: 19648965
  97. The protein and gene expression of the M3 receptor in the prostatic carcinoma group were higher than that of benign prostatic hyperplasia group and normal prostate group. PMID: 19668880
  98. m3 receptor-expressing HeLa cells are a valuable system for studying IP(3) receptor ERAD, and suggest that the SPFH1/2 complex is a factor that selectively mediates the ERAD of activated IP(3) receptors. PMID: 19751772

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Involvement in disease Prune belly syndrome (PBS)
Subcellular Location Cell membrane, Multi-pass membrane protein, Cell junction, synapse, postsynaptic cell membrane, Multi-pass membrane protein, Basolateral cell membrane, Multi-pass membrane protein
Protein Families G-protein coupled receptor 1 family, Muscarinic acetylcholine receptor subfamily, CHRM3 sub-subfamily
Database Links

HGNC: 1952

OMIM: 100100

KEGG: hsa:1131

STRING: 9606.ENSP00000255380

UniGene: Hs.155736

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