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Acetylthiocholine iodide

Acetylthiocholine iodide is a synthetic compound used as a cholinergic agonist and as a substrate for the enzyme acetylcholinesterase.
It is commonly employed in research and diagnostic applications, such as assessing cholinergic function and measuring acetylcholinesterase activity.
This chemical compound can be a useful tool in the study of the cholinergic system and related pathways.
Researchers can utilize PubCompare.ai to discover optimal protocols and methods for working with acetylthiocholine iodide and streamline their experimental approches.

Most cited protocols related to «Acetylthiocholine iodide»

Comprehensive Antioxidant and Enzyme Inhibitory Assays

Antioxidant (DPPH and ABTS radical scavenging, reducing power (CUPRAC and FRAP), phosphomolybdenum, and metal chelating (ferrozine method)) and enzyme inhibitory activities [cholinesterase (ChE) Elmann’s method], tyrosinase (dopachrome method), α-amylase (iodine/potassium iodide method), and α -glucosidase (chromogenic PNPG method)) were determined using the methods previously described by Zengin et al. (2014) (link) and Dezsi et al. (2015) (link).
For the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging assay: Sample solution (1 mg/mL; 1 mL) was added to 4 mL of a 0.004% methanol solution of DPPH. The sample absorbance was read at 517 nm after a 30 min incubation at room temperature in the dark. DPPH radical scavenging activity was expressed as millimoles of trolox equivalents (mg TE/g extract).
For ABTS (2,2′-azino-bis(3-ethylbenzothiazoline) 6-sulfonic acid) radical scavenging assay: Briefly, ABTS+ was produced directly by reacting 7 mM ABTS solution with 2.45 mM potassium persulfate and allowing the mixture to stand for 12–16 in the dark at room temperature. Prior to beginning the assay, ABTS solution was diluted with methanol to an absorbance of 0.700 ± 0.02 at 734 nm. Sample solution (1 mg/mL; 1 mL) was added to ABTS solution (2 mL) and mixed. The sample absorbance was read at 734 nm after a 30 min incubation at room temperature. The ABTS radical scavenging activity was expressed as millimoles of trolox equivalents (mmol TE/g extract) (Mocan et al., 2016a (link)).
For CUPRAC (cupric ion reducing activity) activity assay: Sample solution (1 mg/mL; 0.5 mL) was added to premixed reaction mixture containing CuCl₂ (1 mL, 10 mM), neocuproine (1 mL, 7.5 mM) and NH₄Ac buffer (1 mL, 1 M, pH 7.0). Similarly, a blank was prepared by adding sample solution (0.5 mL) to premixed reaction mixture (3 mL) without CuCl₂. Then, the sample and blank absorbances were read at 450 nm after a 30 min incubation at room temperature. The absorbance of the blank was subtracted from that of the sample. CUPRAC activity was expressed as milligrams of trolox equivalents (mg TE/g extract).
For FRAP (ferric reducing antioxidant power) activity assay: Sample solution (1 mg/mL; 0.1 mL) was added to premixed FRAP reagent (2 mL) containing acetate buffer (0.3 M, pH 3.6), 2,4,6-tris(2-pyridyl)-S-triazine (TPTZ) (10 mM) in 40 mM HCl and ferric chloride (20 mM) in a ratio of 10:1:1 (v/v/v). Then, the sample absorbance was read at 593 nm after a 30 min incubation at room temperature. FRAP activity was expressed as milligrams of trolox equivalents (mg TE/g extract).
For phosphomolybdenum method: Sample solution (1 mg/mL; 0.3 mL) was combined with 3 mL of reagent solution (0.6 M sulfuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate). The sample absorbance was read at 695 nm after a 90 min incubation at 95°C. The total antioxidant capacity was expressed as millimoles of trolox equivalents (mmol TE/g extract) (Mocan et al., 2016c (link)).
For metal chelating activity assay: Briefly, sample solution (1 mg/mL; 2 mL) was added to FeCl₂ solution (0.05 mL, 2 mM). The reaction was initiated by the addition of 5 mM ferrozine (0.2 mL). Similarly, a blank was prepared by adding sample solution (2 mL) to FeCl₂ solution (0.05 mL, 2 mM) and water (0.2 mL) without ferrozine. Then, the sample and blank absorbances were read at 562 nm after 10 min incubation at room temperature. The absorbance of the blank was sub-tracted from that of the sample. The metal chelating activity was expressed as milligrams of EDTA (disodium edetate) equivalents (mg EDTAE/g extract).
For ChE inhibitory activity assay: Sample solution (1 mg/mL; 50 μL) was mixed with DTNB (5,5-dithio-bis(2-nitrobenzoic) acid, Sigma, St. Louis, MO, United States) (125 μL) and AChE [acetylcholines-terase (Electric ell AChE, Type-VI-S, EC 3.1.1.7, Sigma)], or BChE [BChE (horse serum BChE, EC 3.1.1.8, Sigma)] solution (25 μL) in Tris–HCl buffer (pH 8.0) in a 96-well microplate and incubated for 15 min at 25°C. The reaction was then initiated with the addition of acetylthiocholine iodide (ATCI, Sigma) or butyrylthiocholine chloride (BTCl, Sigma) (25 μL). Similarly, a blank was prepared by adding sample solution to all reaction reagents without enzyme (AChE or BChE) solution. The sample and blank absorbances were read at 405 nm after 10 min incubation at 25°C. The absorbance of the blank was subtracted from that of the sample and the cholinesterase inhibitory activity was expressed as galanthamine equivalents (mgGALAE/g extract) (Mocan et al., 2016b (link)).
For Tyrosinase inhibitory activity assay: Sample solution (1 mg/mL; 25 μL) was mixed with tyrosinase solution (40 μL, Sigma) and phosphate buffer (100 μL, pH 6.8) in a 96-well microplate and incubated for 15 min at 25°C. The reaction was then initiated with the addition of L-DOPA (40 μL, Sigma). Similarly, a blank was prepared by adding sample solution to all reaction reagents without enzyme (tyrosinase) solution. The sample and blank absorbances were read at 492 nm after a 10 min incubation at 25°C. The absorbance of the blank was subtracted from that of the sample and the tyrosinase inhibitory activity was expressed as kojic acid equivalents (mgKAE/g extract) (Mocan et al., 2017 (link)).
For α-amylase inhibitory activity assay: Sample solution (1 mg/mL; 25 μL) was mixed with α-amylase solution (ex-porcine pancreas, EC 3.2.1.1, Sigma) (50 μL) in phosphate buffer (pH 6.9 with 6 mM sodium chloride) in a 96-well microplate and incubated for 10 min at 37°C. After pre-incubation, the reaction was initiated with the addition of starch solution (50 μL, 0.05%). Similarly, a blank was prepared by adding sample solution to all reaction reagents without enzyme (α-amylase) solution. The reaction mixture was incubated 10 min at 37°C. The reaction was then stopped with the addition of HCl (25 μL, 1 M). This was followed by addition of the iodine-potassium iodide solution (100 μL). The sample and blank absorbances were read at 630 nm. The absorbance of the blank was subtracted from that of the sample and the α-amylase inhibitory activity was expressed as acarbose equivalents (mmol ACE/g extract) (Savran et al., 2016 (link)).
For α-glucosidase inhibitory activity assay: Sample solution (1 mg/mL; 50 μL) was mixed with glutathione (50 μL), α-glucosidase solution (from Saccharomyces cerevisiae, EC 3.2.1.20, Sigma) (50 μL) in phosphate buffer (pH 6.8) and PNPG (4-N-trophenyl-α-D-glucopyranoside, Sigma) (50 μL) in a 96-well microplate and incubated for 15 min at 37°C. Similarly, a blank was prepared by adding sample solution to all reaction reagents without enzyme (α-glucosidase) solution. The reaction was then stopped with the addition of sodium carbonate (50 μL, 0.2 M). The sample and blank absorbances were read at 400 nm. The absorbance of the blank was subtracted from that of the sample and the α-glucosidase inhibitory activity was expressed as acarbose equivalents (mmol ACE/g extract) (Llorent-Martínez et al., 2016 (link)).
All the assays were carried out in triplicate. The results are expressed as mean values and standard deviation (SD). The differences between the different extracts were analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s honestly significant difference post hoc test with α = 0.05. This treatment was carried out using SPSS v. 14.0 program.

Uysal S., Zengin G., Locatelli M., Bahadori M.B., Mocan A., Bellagamba G., De Luca E., Mollica A, & Aktumsek A. (2017). Cytotoxic and Enzyme Inhibitory Potential of Two Potentilla species (P. speciosa L. and P. reptans Willd.) and Their Chemical Composition. Frontiers in Pharmacology, 8, 290.

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Publication 2017

Cytoarchitecture of Mouse Prefrontal Cortex

The cytoarchitecture of the PFC was studied in ten adult, male mice (strain C57BL/6) of similar weight (approximately 20 g). These control mouse brains were kindly donated and immersion fixed by Dr. H. Manji, NIMH, USA. All animal procedures were in strict accordance with the NIH animal care guidelines. The histological processing of these brains was performed at the laboratory of Dr. Rajkowska. The brains were embedded in 12% celloidin, cut into 40-μm serial sections using a sliding microtome and Nissl (1% cresyl violet) stained. Celloidin was chosen as an embedding medium to allow for the preparation of ‘thick’ sections with clear morphology and high contrast of Nissl-stained neurons and glial cells. In these immersion-fixed brains, any spots showing pycnotic reaction were not incorporated in this study.
In addition to these ten mice, four adult male mice (C57BL/6 strain) were stained for dopamine and four adult male mice for AChE, myelin, and immunohistochemically for SMI, PV and CB. For each staining, a different set of sections with several consecutive sections stained with Nissl at HBMU’s laboratory was used. The antibodies applied were the dopamine (DA) antibody (Geffard et al. 1984 (link)), SMI-32 antibody (Sternberger Monoclonals Inc., Baltimore, MD, USA: monoclonal antibody to one epitope of non-phosphorylated tau neurofilaments, lot number 11), SMI-311antibody (pan-neuronal neurofilament marker cocktail of several monoclonal antibodies for several epitopes of non-phosphorylated tau protein, Sternberger Monoclonals Inc., Baltimore, MD, USA: lot number 9) (SMI antibodies are presently distributed through Covance Research Products, USA), monoclonal anti-CB D-28K antibody (Sigma, St. Louis, MO, USA: product number C-9848, clone number CB-955, lot number 015K4826), and monoclonal anti-PV antibody (Sigma, St. Louis, MO, USA: product number P-3171, clone number PA-235, lot number 026H4824). Mice to be stained for DA were intracardially perfused under deep pentobarbital anesthesia (1 ml/kg body weight, i.p.), with saline followed by fixative. For DA staining, the fixative was 5% glutaraldehyde in 0.05 M acetate buffer at pH 4.0. After perfusion, the brains were immersed in 0.05 Tris containing 1% sodium disulfite (Na₂S₂O₅) at pH 7.2 (De Brabander et al. 1992 (link)). Mouse PFC was sectioned at 40 μm by a vibratome. These sections were stained overnight in a cold room at 4°C using the polyclonal primary antibody sensitive to DA that was raised in the Netherlands Institute for Brain Research (NIBR) (Geffard et al. 1984 (link)), the specificity of which had been demonstrated previously (Kalsbeek et al. 1990 (link)). DA antiserum was diluted 1:2,000 in 0.05 M Tris containing 1% Na₂S₂O₅ and 0.5% Triton X-100, pH 7.2. After overnight incubation, the sections were washed three times with Tris-buffered saline (TBS) and subsequently incubated in the secondary antibody goat–antirabbit, also raised in NIBR at 1:100 for 1 h. After having been rinsed 3× in TBS, it was incubated in the tertiary antibody, peroxidase–antiperoxidase, at 1:1,000 for 60 min. Both the secondary and the tertiary antibodies were diluted in TBS with 0.5% gelatine and 0.5% Triton X-100. For visualization, the sections were transferred into 0.05% diaminobenzidine (DAB; Sigma) with 0.5% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring the sections to TBS (3 × 10 min), then the sections were mounted on slides, air dried, washed, dehydrated and coverslipped.
Mice to be stained with anti-PV, anti-CB and SMI-32 and SMI-311 were fixed with 4% formaldehyde solution in 0.1 M phosphate buffer at pH 7.6. Mouse PFC was sectioned at 40 μm by a vibratome. To prevent endogenous peroxidase activity, free-floating sections were pretreated for 30 min in a Tris-buffered saline (TBS) solution containing 3% hydrogen peroxide and 0.2% Triton X-100. To prevent non-specific antibody staining, these sections were placed in a milk solution (TBS containing 5% nonfat dry milk and 0.2% Triton X-100) for 1 h. Incubation of the primary antibody, directly after the milk step was carried out overnight in a cold room at 4°C. The primary antibodies were diluted in the above-mentioned milk solution: SMI-32 and SMI-311 at 1:1,000, PV antibody at 1:1,000, and CB antibody at 1:250. For the monoclonal SMI-32, SMI-311, PV and CB antibodies, raised in mice, we used peroxidase-conjugated rabbit–antimouse (1:100 in 5% milk solution with 0.2% Triton X-100) as a secondary antibody. Visualization took place in 0.05% diaminobenzidine enhanced with 0.2% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring these sections to TBS (3 × 10 min), after which the sections were rinsed in distilled water, mounted on slides, air dried, washed, dehydrated and coverslipped. Control sections that were incubated according to the same procedure as described above, omitting the primary antibody, were all negative. All sections were cut coronally, because the coronal plane offers in general the best view to differentiate between the subareas of the rodent PFC (Uylings et al. 2003 (link); Van de Werd and Uylings 2008 (link)).
Sections were processed for AChE staining according to the protocol described by Cavada et al. (1995 (link)). The sections were incubated overnight in a solution of cupric sulfate and acetate buffer at pH 5 to which acetylthiocholine iodide and ethopropazine were added just before the start of incubation. After rinsing, the sections were developed in a sodium sulfide solution until a light brown color appeared and subsequently intensified to a dark brown color in a silver nitrate solution. Finally, the sections were differentiated after rinsing in a thiosulfate solution, dehydrated and mounted. In all steps, the solutions and sections were shaken constantly. The myelin was stained with silver by physical development according to Gallyas (1979 (link)). The sections were first placed in 100% ethanol and then immersed in a 2:1 solution of pyridine and acetic acid for 30 min. After rinsing, they were placed in an ammonium silver nitrate solution and after rinsing with 0.5% acetic acid, the sections were immersed in the optimal physical developer solution at room temperature (Gallyas 1979 (link)) until they showed good stain intensity under the microscope. Then the development of the staining was stopped in 0.5% acetic acid and the sections were dehydrated and mounted with Histomount. The sections were studied at intervals of 80–160 μm, and examined under a light microscope at a 63× magnification.

Van De Werd H.J., Rajkowska G., Evers P, & Uylings H.B. (2010). Cytoarchitectonic and chemoarchitectonic characterization of the prefrontal cortical areas in the mouse. Brain Structure & Function, 214(4), 339-353.

Publication 2010

Inhibition of Acetylcholinesterase and Antioxidant Activity

AChE from Electrophorus electricus (CAS: 9000-81-1) was obtained from Sigma Aldrich, St. Loius, MO, United States, while BChE was derived from equine serum (9001-08-5) and was purchased from Sigma–Aldrich GmbH, Germany. Acetylthiocholine iodide (CAS1866-15-5) and butyrylthiocholine iodide (CAS 2494-56-6) were purchased from Sigma–Aldrich United Kingdom and Sigma–Aldrich Switzerland respectively. 5,5-Dithio-bis-nitrobenzoic acid (DTNB) (CAS 69-78-3) (Sigma–Aldrich GmbH, Germany) and galanthamine HBr Lycoris Sp. (CAS: 1953-04-4) (Sigma–Aldrich, France) were used in enzymes studies. Antioxidant reagents including DPPH (CAS: 1898-66-4) ABTS (CAS: 30931-67-0) were purchased from Sigma Aldrich St. Loius, MO, United States. H₂O₂ (batch no: A040) was obtained from Rehmat pharma Lahore, Pakistan. Potassium peroxodisulfate (LOT NO: 51240) was obtained from Labor chemikalien GmbH & Co KGD-30926 Seelze. For genotyping of transgenic animals, GF-1 tissue DNA extraction kit (Cat:GF-TD-100, Vivantis), agarose (Invitrogen CAT:75510-011, Carlsbad, CA, United States), boric acid (Serva CAT 15165, Germany), DNA Ladder (Serva CAT:15165, Germany), EDTA (Invitrogen CAT:75576-028, Carlsbad, CA, United States), ethanol (Merck CAT:26225745, Germany), ethidium bromide (Sigma CAT:E7637, United States), MgCl₂ (Invitrogen CAT:AM9530G, Carlsbad, CA, United States), DNTPs (Promega CAT:U1515, United States), Taq polymerase (Thermo Scientific CAT: EP0402, United States), PCR primers (Thermo Scientific CAT:OIMR3610 F, OIMR3611 R), PCR grade distilled water (Thermo Scientific CAT: R0581), sucrose (Invitrogen CAT: 15503-022, Carlsbad, CA, United States), Tris EDTA solution (50X), 2XPCR Master mix (Fermentas CAT: K0171, EU), NaCl (Invitrogen CAT: 24740-011, Carlsbad, CA, United States) and tris (Invitrogen CAT: 15504-020, Carlsbad, CA, United States) were purchased from authorized dealers in Pakistan. Solvents and buffer salts used were of extra pure quality.

Ayaz M., Junaid M., Ullah F., Subhan F., Sadiq A., Ali G., Ovais M., Shahid M., Ahmad A., Wadood A., El-Shazly M., Ahmad N, & Ahmad S. (2017). Anti-Alzheimer’s Studies on β-Sitosterol Isolated from Polygonum hydropiper L.. Frontiers in Pharmacology, 8, 697.

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Publication 2017

2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid acetylthiocholine iodide Animals, Transgenic Antioxidants boric acid Buffers Butyrylthiocholine Edetic Acid Electric Eel Enzymes Equus caballus Ethanol Ethidium Bromide Galanthamine Hydrobromide Iodides Lycoris Magnesium Chloride Nitrobenzoic Acids Obstetric Labor Oligonucleotide Primers Pain Peroxide, Hydrogen potassium persulfate Promega Salts Sepharose Serum Sodium Chloride Solvents Sucrose Taq Polymerase Tissues Tromethamine

Cytoarchitecture of Murine Prefrontal Cortex

The cytoarchitecture of the prefrontal cortex was studied in ten adult, male mice (strain C57BL/6) of similar weight (approximately 20g). These control mouse brains were kindly donated and immersed fixed by Dr. H. Manji, NIMH, USA. All animal procedures were in a strict accordance with the NIH animal care guidelines. The histological processing of these brains was performed at the laboratory of Dr. Rajkowska. The brains were embedded in 12% celloidin, cut into 40μm serial sections using a sliding microtome and Nissl (1% cresyl violet) stained. Celloidin was chosen as an embedding medium to allow for the preparation of ‘thick’ sections with clear morphology and high contrast of Nissl stained neurons and glial cells. In these immersion-fixed brains, any spots showing pycnotic reaction were not incorporated in this study.
In addition to these ten mice, four adult male mice (C57BL/6 strain) have been stained for dopamine and four adult male mice for acetylcholinesterase (AChE), myelin, and immunohistochemically for SMI, parvalbumin and calbindin: per staining a different set of sections with several consecutive sections stained with Nissl at HBMU's laboratory. The antibodies applied are the dopamine (DA) antibody (Geffard et al., 1984 (link)), SMI-32 antibody (Sternberger Monoclonals Inc., Baltimore, MD, USA: monoclonal antibody to one epitope of non-phosphorylated tau neurofilaments, Lot Number: 11), SMI-311antibody (Pan-Neuronal Neurofilament Marker cocktail of several monoclonal antibodies for several epitopes of non-phosphorylated tau protein, Sternberger Monoclonals Inc., Baltimore, MD, USA: Lot Number 9) [SMI antibodies are presently distributed through Covance Research Products, USA], monoclonal anticalbindin D-28K antibody (Sigma, St. Louis, Mo., USA: Product Number: C-9848, Clone Number: CB-955, Lot Number: 015K4826), and monoclonal anti-parvalbumin antibody (Sigma, St. Louis, Mo., USA: Product Number: P-3171, Clone Number: PA-235, Lot Number: 026H4824). Mice to be stained for DA were intracardially perfused under deep pentobarbital anaesthesia (1 ml/kg body weight, i.p.), with saline followed by fixative. For DA staining the fixative was 5% glutaraldehyde in 0.05 M acetate buffer at pH 4.0. After perfusion the brains were immersed in 0.05 Tris containing 1% Sodium disulfite (Na₂S₂O₅) at pH 7.2 (De Brabander et al. 1992 (link)). Mouse prefrontal cortex was sectioned at 40 μm by a vibratome. These sections were stained overnight in a cold room at 4° C. using the polyclonal primary antibody sensitive for DA raised in the Netherlands Institute for Brain Research (NIBR) (Geffard et al. 1984 (link)) of which the specificity has been demonstrated previously (Kalsbeek et al. 1990 (link)). DA antiserum was diluted 1:2,000 in 0.05 M Tris containing 1% Na₂S₂O₅ and 0.5% Triton X-100, pH 7.2. After overnight incubation the sections were washed three times with Tris buffered saline (TBS) and subsequently incubated in the secondary antibody goat-antirabbit, also raised in NIBR, at 1:100 for 1 h. and, after having been rinsed 3x in TBS, incubated in the tertiary antibody, peroxidase-antiperoxidase, at 1:1,000 for 60 min. Both the secondary and the tertiary antibodies were diluted in TBS with 0.5% gelatine and 0.5% Triton X-100. For visualization the sections were transferred into 0.05% diaminobenzidine 9(DAB; Sigma) with 0.5% nickel ammonium sulphate. The reaction was stopped after a few minutes by transferring the sections to TBS (3x 10 min), then the sections were mounted on slides, air dried, washed, dehydrated and coverslipped. Mice to be stained with antiparvalbumin, anticalbindin and SMI-32 and SMI-311 were fixed with 4% formaldehyde solution in 0.1 M phosphate buffer at pH 7.6. Mouse prefrontal cortex was sectioned at 40 μm by a vibratome. To prevent endogenous peroxidase activity free-floating sections were pretreated for 30 min in a Tris-buffered saline (TBS) solution containing 3% hydrogen peroxide and 0.2% Triton X-100. To prevent non-specific antibody staining, these sections were placed in a milk solution (TBS containing 5% nonfat dry milk and 0.2% Triton X-100) for 1 h. Incubation of the primary antibody, directly after the milk step, was carried out overnight in a cold room at 4%. The primary antibodies were diluted in above-mentioned milk solution: SMI-32 and SMI-311 at 1:1,000, parvalbumin antibody at 1:1,000, and calbindin antibody at 1:250. For the monoclonal SMI-32, SMI-311, parvalbumin and calbindin antibodies, raised in mice, we used peroxidase-conjugated rabbit-antimouse (1:100 in 5% milk solution with 0.2% Triton X-100) as a secondary antibody. Visualization took place in 0.05% diaminobenzidine enhanced with 0.2% nickel ammonium sulphate. The reaction was stopped after a few minutes by transferring these sections to TBS (3x 10 min), after which the sections were rinsed in distilled water, mounted on slides, air dried, washed, dehydrated and coverslipped. Control sections incubated according to the same procedure as described above, but omitting the primary antibody were all negative. All sections have been cut coronally, because the coronal plane offers in general the best view to differentiate between the subareas of the rodent prefrontal cortex (Uylings et al. 2003 (link); Van de Werd and Uylings 2008 (link)).
The sections were studied at intervals of 80-160 μm, and were examined under the light-microscope at a 63x magnification.
Sections have been processed for acetylcholinesterase staining according to protocol described by Cavada et al. (1995) (link). The sections were incubated overnight in a solution of cupric sulphate and acetate buffer at pH 5 to which acetylthiocholine iodide and ethopropazine were added just before the start of the incubation. After rinsing the sections were developed in a sodium sulphide solution until a light brown color appeared and subsequently intensified in a silver nitrate solution until a dark brown color. Finally, the sections are differentiated after rinsing in a thiosulphate solution, dehydrated and mounted. In all steps the solutions and sections were shaken constantly. The myelin was stained with silver by means of physical development according to Gallyas (1979) (link). The sections were first put into 100% ethanol, and then immersed in a 2:1 solution of pyridine and acetic acid for 30 minutes. After rinsing they were brought into an ammonium-silver-nitrate solution and after rinsing them in 0.5% acetic acid, the sections were immersed in the optimal physical developer solution for room temperature (Gallyas, 1979 (link)) until the sections showed a good stain-intensity, checked under microscope. Then the development of the staining was stopped in 0.5% acetic acid and the sections were dehydrated and mounted with Histomount.

Publication 2010

Histochemical Delineation of Amygdalar Nuclei

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Publication 2016

Most recents protocols related to «Acetylthiocholine iodide»

Flavonoid Inhibition of BChE/AChE Activities

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Publication 2024

Measuring Acetylcholinesterase Activity

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A reaction mixture consisting of 50 µl of tissue homogenate, 3 ml of 0.01 M sodium phosphate buffer (pH 8.0), 100 µl of acetylthiocholine iodide, and 100µl5,5, dithiobis-(2-nitro benzoic acid) was taken. AChE activity was measured using molar extinction co-e cient 1.36×10 4 M - 1 cm - 1 at 412 nm and measured as nmol of acetylthiocholine iodide hydrolyzed/min/mg protein[26].

Dhaliwal N., Dhaliwal J, & Chopra K. (2024). 7, 8-dihydroxyflavone Ameliorates Cholinergic Dysfunction, Inflammation, Oxidative Stress, and Apoptosis in a Rat Model of Vascular Dementia. Neurochemical research, 49(5).

Publication 2024

Cholinesterase Inhibition Assay Protocol

To evaluate the therapeutic effectiveness of the compounds as inhibitors of cholinesterases, the reported procedures were followed (Elman's protocol) [31 ]. Enzymes acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8) were purchased from local market.
The acetylcholinesterase inhibition assay is based on the principle of the hydrolytic breakdown of substrate; acetylthiocholine iodide by acetylcholinesterase enzyme. The AChE acts on acetylthiocholine to thiocholine which then react with DTNB that's leads towards the formations of anion (5-thio-2-nitrobenzoate) a yellow colour product which is then assessed via UV visible spectrophotometer at 405 nm [32 ].
The AChE, Ellman's Reagent (DTNB) and acetylthiocholine iodide (ATChI, substrate), and the test compound solutions were made in different concentrations ranges from (0.01–1.0 μM) were incubated per detail given in the Ellman's protocol. Galantamine was employed as the reference drug [33 (link),34 ]. IC₅₀ of each Schiff bases were calculated using Microsoft Excel 2013 software.
In similar fashion following BChE, Ellman's Reagent (DTNB) and butyryl thiocholine iodide (BTChI, substrate), and the test compound Schiff bases solutions were made in different concentrations ranges from (0.01–1.0 μM) were incubated following the standard protocol. The absorbance at 405 nm was measured and the data were analysed in triplicate. Galantamine was employed as standard drug [33 (link),34 ].

Gul Q., Karim N., Shoaib M., Zahoor M., Rahman M.U., Bilal H., Ullah R, & Alotaibi A. (2024). Vanillin derivatives as antiamnesic agents in scopolamine-induced memory impairment in mice. Heliyon, 10(4), e26657.

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Publication 2024

Acetylcholinesterase Activity Assay

Not available on PMC !

The acetylcholinesterase activities of the extracts of M. myrtifolia were measured with the AChE activity of Electrophorus electricus (electric eels) spectrophotometrically using the Ellman Method [23] (link). The substrate used in the study was acetylthiocholine iodide (ATC). The reactions were carried out at 25°C, in 100 mM Tris HCl (pH 8.0) buffer. To start each enzymatic reaction approximately 0.05 U/ml AChE was added to the reaction mixture. The breakdown of acetylthiocholine was detected with a UV spectrophotometer (Carry 60 Single Beam spectrophotometer, Agilent Technologies, USA) over an elevation in absorbance at 412 nm.

Yücel Y.Y., & Nath E.Ö. (2024). MICROMERIA MYRTIFOLIA BOISS. & HOHEN’İN BİYOLOJİK AKTİVİTELERİNİN BELİRLENMESİ. Ankara Universitesi Eczacilik Fakultesi Dergisi, 48(2), 2-2.

Publication 2024

Acetylcholinesterase Activity Assay

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This assay was based on an earlier method reported by Ellman et al. [37] (link) based on the ability of the acetylcholinesterase enzyme to hydrolyse acetylthiocholine producing a thiocholine that reacts with DTNB which gives off a sharp yellow colour. Brie y, 2.85ml of the buffer was placed on the test tube followed by 50 µL of DTNB, 50 µL sample and 20 µL of 78 mM acetylthiocholine iodide and the rate of absorbance increase was monitored over 5 minutes at 412 nm using a spectrophotometer and was expressed as µmoles of substrate hydrolysed/min/mg protein.

Ichipi-Ifukor P.C., Asagba S.O, & Achuba F.I. (2024). Co-exposure to Aluminium and Cadmium Mediates Postpartum Maternal Variation in Brain Architecture and Behaviour of Mice; Involvement of Oxido-nitrergic and Cholinergic Mechanisms : Postpartum effects of Aluminium and Cadmium co-exposure in pregnancy. Biological trace element research.

Publication 2024

Top products related to «Acetylthiocholine iodide»

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Acetylthiocholine iodide is a chemical compound used as a substrate in enzymatic assays. It is commonly employed in the measurement of the activity of the enzyme acetylcholinesterase.

5 5 dithiobis 2 nitrobenzoic acid dtnb by Merck Group

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5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) is a chemical compound used in various laboratory applications. It is a water-soluble, yellow-colored reagent that is commonly employed for the determination of thiol groups in proteins and other biological samples.

5 5 dithiobis 2 nitrobenzoic acid by Merck Group

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Acetylcholinesterase by Merck Group

Sourced in United States, Germany, Italy, Chile, Spain

Acetylcholinesterase is an enzyme that catalyzes the breakdown of the neurotransmitter acetylcholine in the synaptic cleft. It is an important component in the regulation of nerve impulse transmission.

Bovine serum albumin (bsa) by Merck Group

Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico

Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.

Gallic acid by Merck Group

Sourced in United States, Germany, Italy, Spain, France, India, China, Poland, Australia, United Kingdom, Sao Tome and Principe, Brazil, Chile, Ireland, Canada, Singapore, Switzerland, Malaysia, Portugal, Mexico, Hungary, New Zealand, Belgium, Czechia, Macao, Hong Kong, Sweden, Argentina, Cameroon, Japan, Slovakia, Serbia

Gallic acid is a naturally occurring organic compound that can be used as a laboratory reagent. It is a white to light tan crystalline solid with the chemical formula C6H2(OH)3COOH. Gallic acid is commonly used in various analytical and research applications.

2 2 diphenyl 1 picrylhydrazyl (dpph) by Merck Group

Sourced in United States, Germany, Italy, India, China, Spain, Poland, France, United Kingdom, Australia, Brazil, Singapore, Switzerland, Hungary, Mexico, Japan, Denmark, Sao Tome and Principe, Chile, Malaysia, Argentina, Belgium, Cameroon, Canada, Ireland, Portugal, Israel, Romania, Czechia, Macao, Indonesia

DPPH is a chemical compound used as a free radical scavenger in various analytical techniques. It is commonly used to assess the antioxidant activity of substances. The core function of DPPH is to serve as a stable free radical that can be reduced, resulting in a color change that can be measured spectrophotometrically.

Butyrylthiocholine iodide by Merck Group

Sourced in United States, Germany, United Kingdom, Italy

Butyrylthiocholine iodide is a chemical compound used in various laboratory applications. It functions as a substrate for the enzyme butyrylcholinesterase, which is commonly used in the analysis and detection of this enzyme.

Quercetin by Merck Group

Sourced in United States, Germany, Italy, India, Spain, United Kingdom, France, Poland, China, Sao Tome and Principe, Australia, Brazil, Macao, Switzerland, Canada, Chile, Japan, Singapore, Ireland, Mexico, Portugal, Sweden, Malaysia, Hungary

Quercetin is a natural compound found in various plants, including fruits and vegetables. It is a type of flavonoid with antioxidant properties. Quercetin is often used as a reference standard in analytical procedures and research applications.

Ascorbic acid by Merck Group

Sourced in United States, Germany, United Kingdom, France, Italy, India, China, Sao Tome and Principe, Canada, Spain, Macao, Australia, Japan, Portugal, Hungary, Brazil, Singapore, Switzerland, Poland, Belgium, Ireland, Austria, Mexico, Israel, Sweden, Indonesia, Chile, Saudi Arabia, New Zealand, Gabon, Czechia, Malaysia

Ascorbic acid is a chemical compound commonly known as Vitamin C. It is a water-soluble vitamin that plays a role in various physiological processes. As a laboratory product, ascorbic acid is used as a reducing agent, antioxidant, and pH regulator in various applications.

What are some common applications of Acetylthiocholine iodide?

Acetylthiocholine iodide is a widely used chemical compound in research and diagnostic applications. It is commonly employed as a cholinergic agonist, meaning it can activate cholinergic receptors, and as a substrate for the enzyme acetylcholinesterase. Researchers often use acetylthiocholine iodide to assess cholinergic function and measure acetylcholinesterase activity, which can provide insights into the cholinergic system and related pathways.

What are some key considerations when working with Acetylthiocholine iodide?

When using Acetylthiocholine iodide, researchers should be aware of a few important factors. Firstly, the compound can be sensitive to hydrolysis, so it's crucial to store it properly and use it within the recommended timeframe to ensure optimal activity. Additionally, the concentration and purity of the compound can impact experimental results, so researchers should carefully consider these variables when designing their studies.

How can PubCompare.ai help researchers work with Acetylthiocholine iodide more effectively?

PubCompare.ai is a powerful AI-driven platform that can assist researchers in optimizing their protocols and methods when working with Acetylthiocholine iodide. The platform allows you to efficiently screen protocol literature, leveraging AI to pinpoint critical insights that can help you identify the most effective protocols for your specific research goals. By highlighting key differences in protocol effectiveness, PubCompare.ai enables you to choose the best option for reproducibility and accuracy, streamlining your research with Acetylthiocholine iodide.

Are there any variations or types of Acetylthiocholine iodide that researchers should be aware of?

Yes, there are a few different variations of Acetylthiocholine iodide that researchers may encounter. In addition to the standard form, there are also radioactively labeled versions of the compound, such as [3H]Acetylthiocholine iodide and [14C]Acetylthiocholine iodide, which can be used in specialized studies involving radioactivity. Researchers should carefully consider the specific needs of their experiments when selecting the appropriate form of Acetylthiocholine iodite.

More about "Acetylthiocholine iodide"

Acetylthiocholine iodide, also known as Acetyl-β-methylthiocholine iodide (AMTI) or ATCh, is a synthetic chemical compound commonly used in research and diagnostic applications.
It is a cholinergic agonist, meaning it can mimic the effects of the neurotransmitter acetylcholine, and is also a substrate for the enzyme acetylcholinesterase (AChE).
The primary uses of acetylthiocholine iodide include assessing cholinergic function and measuring AChE activity.
Researchers often employ this chemical compound to study the cholinergic system and related pathways, which are involved in various physiological processes, such as neurotransmission, muscle contraction, and cognitive function.
In addition to acetylthiocholine iodide, other related compounds like 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), also known as Ellman's reagent, are commonly used in conjunction with acetylthiocholine iodide to measure AChE activity.
DTNB reacts with the free sulfhydryl groups released during the enzymatic hydrolysis of acetylthiocholine iodide, allowing for the quantification of AChE activity.
Furthermore, acetylthiocholine iodide can be used in combination with other substances, such as bovine serum albumin, gallic acid, DPPH, butyrylthiocholine iodide, quercetin, and ascorbic acid, in various experimental setups and assays.
These additional compounds may serve as cofactors, substrates, or inhibitors, depending on the specific research objectives.
Researchers can utilize PubCompare.ai, an AI-powered research platform, to discover optimal protocols and methods for working with acetylthiocholine iodide and streamline their experimental approaches.
PubCompare.ai helps researchers identify the most accurate and reproducible protocols from literature, preprints, and patents, enabling data-driven decision-making and enhancing the efficiency of their studies.