Acetylcholinesterase Assay Kit

Produktnummer ABIN1000245

Produktdetails

Target: Acetylcholinesterase (AChE)

Reaktivität: Chemical

Applikation: Activity Assay (AcA)

Proben: Blood, Plasma, Serum

Spezifität: 10 U/L

Produktmerkmale: Sensitive and accurate. Detection range 10 to 600 U/L AChE activity in 96-well plate assay.
Convenient. The procedure involves adding a single working reagent, and reading the optical density at 2 min and 10 min at room temperature.
High-throughput. Can be readily automated as a high-throughput 96- well plate assay for thousands of samples per day.

Bestandteile: Assay Buffer (pH 7.5): 30 mL. Reagent: 240 mg. Calibrator: 4 mL (equivalent to 200 U/L).

Benötigtes Material: Pipeting (multi-channel) devices. Clear-bottom 96-well plates (e.g. Corning Costar) and plate reader.

Anwendungsinformationen

Applikationshinweise: Direct assays of acetylcholinesterase activity in blood, serum, plasma, and other biological samples. Evaluation of acetylcholinesterase inhibitors.

Kommentare:

1. This assay is based on an enzyme-catalyzed kinetic reaction. Addition of Working Reagent should be quick and mixing should be brief but thorough. Use of a multi-channel pipettor is recommended.
2. For assays in standard 1 mL cuvet, use 1 mL water and 1 mL Calibrator, 50 μL sample + 950 μL Working Reagent.

Protokoll: 1. Calibrator: transfer 200 µL water and 200 µL calibrator separately into wells of a clear bottom 96-well plate. Samples: add 10 µL sample per well in separate wells.
2. Reaction: transfer 190 µL freshly prepared Working Reagent to all sample wells and tap plate briefly to mix. Read OD412nm at 2 min and at 10 min in a plate reader.

Aufbereitung der Reagenzien:

The Working Reagent should be prepared freshly and used within 30 min. Each reaction well requires 2 mg reagent. Calculate the amount of reagent needed and weigh this amount (mg) in a centrifuge tube. Add 200 µL Assay Buffer per 2 mg reagent. Vortex to dissolve.

Aufbereitung der Proben:

Blood samples should be diluted 40-fold in the Assay Buffer, e.g. accurately pipet 5 µL blood and mix thoroughly with 195 µL Assay Buffer. Tissue or cell lysates are prepared by brief sonication or homogenization in 0.1M phosphate buffer (pH
7.5), followed by centrifugation at 14,000 rpm for 5 min. Use supernatant for assay. Ideally samples should be assayed fresh. If this is not possible, refrigerate samples and assay them within 24 hours.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -20 °C

Referenzen

Mineur, Obayemi, Wigestrand, Fote, Calarco, Li, Picciotto: "Cholinergic signaling in the hippocampus regulates social stress resilience and anxiety- and depression-like behavior." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 110, Issue 9, pp. 3573-8, (2013) (PubMed).

Khokhar, Tyndale: "Rat brain CYP2B-enzymatic activation of chlorpyrifos to the oxon mediates cholinergic neurotoxicity." in: Toxicological sciences : an official journal of the Society of Toxicology, Vol. 126, Issue 2, pp. 325-35, (2012) (PubMed).

Ng, Villuendas, Cokic, Schliamser, Gordon, Koduri, Benefield, Simon, Murthy, Lomasney, Wasserstrom, Goldberger, Aistrup, Arora: "Autonomic remodeling in the left atrium and pulmonary veins in heart failure: creation of a dynamic substrate for atrial fibrillation." in: Circulation. Arrhythmia and electrophysiology, Vol. 4, Issue 3, pp. 388-96, (2011) (PubMed).

Basselin, Nguyen, Chang, Bell, Rapoport: "Acute but not chronic donepezil increases muscarinic receptor-mediated signaling via arachidonic acid in unanesthetized rats." in: Journal of Alzheimer's disease : JAD, Vol. 17, Issue 2, pp. 369-82, (2009) (PubMed).

Al-Akwa, Shaher, Al-Akwa, Aleryani: "Free radicals are present in human serum of Catha edulis Forsk (Khat) abusers." in: Journal of ethnopharmacology, Vol. 125, Issue 3, pp. 471-3, (2009) (PubMed).

Antigendetails

Target: Acetylcholinesterase (AChE)

Andere Bezeichnung: Acetylcholinesterase (AChE Produkte)

Synonyme: ACEE Kit, ARACHE Kit, N-ACHE Kit, YT Kit, Ache Kit, GB14873 Kit, zgc:92550 Kit, ACE Kit, Dsim\\GD20515 Kit, GD20515 Kit, dsim_GLEANR_4292 Kit, ache Kit, mE1a Kit, mE1b Kit, mE1c Kit, mE1c-long Kit, mE1d Kit, mE1d' Kit, mE1e Kit, arache Kit, n-ache Kit, acetylcholinesterase (Cartwright blood group) Kit, acetylcholinesterase 2 Kit, acetylcholinesterase Kit, Acetylcholine esterase Kit, acetylcholinesterase (Cartwright blood group) L homeolog Kit, collagen type I alpha 2 chain Kit, ACHE Kit, AChE-2 Kit, Ache Kit, ache Kit, Dsim\Ace Kit, ache.L Kit, COL1A2 Kit, ACE-1 Kit

Hintergrund: Quantitative determination of acetylcholinesterase activity by colorimetric (412nm) method.
Procedure: 10 min.

Acetylcholinesterase (EC 3.1.1.7, AChE), also known as RBC cholinesterase, is found primarily in the blood and neural synapses. Low serum cholinesterase activity may relate to exposure to insecticides or to one of a number of variant genotypes. AChE catalyzes the hydrolysis of the neurotransmitter acetylcholine into choline and acetic acid, a reaction necessary to allow a cholinergic neuron to return to its resting state after activation. Cholinesterase levels of cells and plasma are used as a guide in establishing safety precautions relative to exposure and contact, as well as a guide in determining the need for workers to be removed from areas of contact with the organic phosphate insecticides. Simple, direct and automation-ready procedures for measuring AChE activity are very desirable. This Acetylcholinesterase Assay is based on an improved Ellman method, in which thiocholine produced by the action of acetylcholinesterase forms a yellow color with 5,5'-dithiobis(2-nitrobenzoic acid). The intensity of the product color, measured at 412 nm, is proportionate to the enzyme activity in the sample.