Reagents and cell culture media were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Life Technologies (Waltham, MA, USA) unless specified otherwise. Commercially obtained compounds had >98% purity. [N-methyl-3H]scopolamine methyl chloride ([3H]NMS) (specific activity 85.4 Ci/mmol) was purchased from PerkinElmer (Waltham, MA, USA). All analytical buffers were prepared in double distilled water (GFL water still 2004). Protease Inhibitor Cocktail powder (P2714-1BTL, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 10 mL water and used as such. Stock solutions of all compounds were prepared in pure DMSO.
N methyl 3h scopolamine methyl chloride 3h nms
Manufactured by PerkinElmer
Sourced in United States
[N-methyl-3H]scopolamine methyl chloride ([3H]NMS) is a radiolabeled compound used in research applications. It serves as a radioligand for the muscarinic acetylcholine receptor.
Automatically generated - may contain errors
Lab products found in correlation
3h rx821002, by PerkinElmer (1 mentions)
Phospho erk1 2, by Santa Cruz Biotechnology (1 mentions)
Calregulin, by Santa Cruz Biotechnology (1 mentions)
Oxo m, by Merck Group (1 mentions)
Antibodies against erk1 2, by Cell Signaling Technology (1 mentions)
Uk14304, by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail powder, by Merck Group (1 mentions)
Pirenzepine, by Bio-Techne (1 mentions)
Calf intestinal alkaline phosphatase, by New England Biolabs (1 mentions)
Telenzepine, by Bio-Techne (1 mentions)
Anti tubulin antibody, by Merck Group (1 mentions)
Qiafilter plasmid maxi kit, by Qiagen (1 mentions)
Wizard plus sv miniprep kit, by Promega (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Merck Group (1 mentions)
Dna restriction endonucleases, by New England Biolabs (1 mentions)
N methylscopolamine, by Merck Group (1 mentions)
T4 ligase, by New England Biolabs (1 mentions)
Nupage mops sds running buffer, by Thermo Fisher Scientific (1 mentions)
Hank s balanced salt solution, by Thermo Fisher Scientific (1 mentions)
4 protocols using n methyl 3h scopolamine methyl chloride 3h nms
1
Radioligand Binding Assay Protocol
2
Biochemical Reagents for Cell Signaling
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Antibodies against GFP, human Rab43, calregulin, phospho-ERK1/2, and β-actin were purchased from Santa Cruz Biotechnology. Antibodies against ERK1/2 were from Cell Signaling Technology. BFA, UK14304, and Oxo-M were obtained from Sigma-Aldrich. The radioligands [3H]-RX821002 (50 Ci/mmol) and [N-methyl-3H]-scopolamine methyl chloride ([3H]-NMS, 80 Ci/mmol) were from PerkinElmer Life Sciences. All other materials were obtained as described elsewhere (63 (link)).
3
Radioligand Binding Assay Protocol
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Reagents and cell culture media were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Life Technologies (Waltham, MA, USA) unless specified otherwise. Commercially obtained compounds had >98% purity. [N-methyl-3H]scopolamine methyl chloride ([3H]NMS) (specific activity 85.4 Ci/mmol) was purchased from PerkinElmer (Waltham, MA, USA). All analytical buffers were prepared in double distilled water (GFL water still 2004). The protease inhibitor cocktail powder (P2714-1BTL, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 10 mL of water and used as such. Stock solutions of all compounds were prepared in pure DMSO.
4
Muscarinic Receptor Binding Assays
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General laboratory chemicals as well as atropine ((RS)-(8-methyl-8-azabicyclo[3.2.1]oct-3-yl) 3-hydroxy-2-phenylpropanoate), N-methylscopolamine (NMS), cytochalasin D, and both anti-tubulin antibody and secondary horseradish peroxidase-conjugated antibody were from Sigma-Aldrich or Fisher Scientific. DNA restriction endonucleases, calf intestinal alkaline phosphatase, T4 DNA polymerase, and T4 ligase were from New England Biolabs (Hitchin, UK). Wizard Plus SV Miniprep kit was from Promega (Southampton, UK). NuPAGE Novex precast 4–12% Bis-Tris gels and NuPAGE MOPS SDS running buffer were from Invitrogen. QIAfilter Plasmid Maxi kit, PCR purification kit, and QIAquick gel extraction kit were from Qiagen (Crawley, UK). Agarose was from Flowgen Biosciences (Nottingham, UK). The anti-GFP antiserum was generated in-house. ECL reagent was purchased from Pierce. [3H]Quinuclidinylbenzilate ([3H]QNB) and [N-methyl-3H]scopolamine methyl chloride ([3H]NMS) were from PerkinElmer Life Sciences. Other muscarinic ligands, specifically pirenzepine and telenzepine were from Tocris (Bristol, UK). Hanks' balanced salt solution was from Life Technologies.
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