This kit will detect acetylcholinesterase and butyrylcholinesterase and will not distinguish between the two.
MAK119
Acetylcholinesterase Activity Assay Kit
sufficient for 100 colorimetric tests
Sinonimo/i:
ACHE Activity Assay, RBC Cholinesterase Activity Assay
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About This Item
Codice UNSPSC:
12161503
NACRES:
NA.84
Prodotti consigliati
impiego
sufficient for 100 colorimetric tests
Metodo di rivelazione
colorimetric
Malattie correlate
neurological disorders
Temperatura di conservazione
room temp
Informazioni sul gene
human ... ACHE(43)
mouse ... ACHE(11423)
Descrizione generale
Acetylcholinesterases (AChEs) are enzymes that hydrolyze the neurotransmitter acetylcholine (ACh) to acetate and choline. AChE is located at the synaptic cleft and functions to terminate synaptic transmission by catalyzing the breakdown of ACh allowing cholinergic neurons to return to a resting state after activation. It is also found in membranes of red blood cells, motor and sensory fibers, muscles, nerves and central and peripheral tissues. Changes in AChE activity may result from exposure to certain insecticides, which act as cholinesterase inhibitors. Inhibitors of AChE are also used to treat certain conditions such as dementia.[1]
Applicazioni
Caratteristiche e vantaggi
Compatible with high-throughput handling systems. Can be adapted for use with cuvettes.
Compatibilità
Suitable for the detection of acetylcholinesterase activity in biological samples such as blood, serum, and plasma.
Principio
This assay is an optimized version of the Ellman method in which thiocholine, produced by AChE, reacts with 5,5′-dithiobis(2-nitrobenzoic acid) to form an colorimetric (412 nm) product, proportional to the AChE activity present. One unit of AChE is the amount of enzyme that catalyzes the production of 1.0 μmole of thiocholine per minute at room temperature at pH 7.5. This kit has a linear range of 10-600 units/L of AChE activity.
Codice della classe di stoccaggio
10 - Combustible liquids
Classe di pericolosità dell'acqua (WGK)
WGK 3
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このキットはAChEに特異的に反応しますか? 血漿中のBuChEには反応しませんか?
1 risposta-
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Is the calibrator in this kit an acetylcholinesterase enzyme, or a colorimetric solution?
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The calibrator is not the enzyme, it is a dye that, when used as directed in the protocol, has an absorbance equivalent to 200U/L under assay conditions.
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Convert the AChE activity (units/L) into a units/mg of tissue value to represent the data?
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To calculate the enzyme activity in U/mg, follow these steps:
AChE Activity = (A412 final – A412 initial) / (A412calibrator – A412 blank) × n × 200 × 0.0002 (U) Here, 0.0002 represents the volume (L) of the well. Specific Activity (U/mg) = AChE Activity (U) / tissue amount (mg)Utile?
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Is it possible to dilute samples by 40x and extend the incubation time to accommodate those that fall below the linear range? Should we adhere to the 10-minute reaction time, or could we consider a longer incubation period to address issues associated with the yellow color continuing to develop when the plate is left out? This consideration arises from the need for a larger change in OD to accurately measure enzymatic activity, particularly due to issues with running out of the Assay Buffer for diluting samples by 40x.
1 risposta-
It is recommended to keep the reaction time at 10 minutes, as the calibrator provided in the kit exhibits the same absorbance as 200 U/L acetylcholinesterase at 10 minutes and does not receive the working reagent. However, to increase the signal of the assay, the customer may increase the sample to working reagent volume. For instance, instead of adding 10 uL of sample to each well, add 20 uL of sample to each well. The plate should still be read at the 2 and 10 minute marks. Additionally, the supplier advises using EDTA as the anticoagulant.
Utile?
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What is the difference between the two Acetylcholinesterase Activity Assay Kits MAK119 and CS0003?
1 risposta-
Product CS0003 and MAK119 are produced by different manufacturers. In addition, CS0003 is sufficient for 200 tests while product MAK119 is sufficient for 100 tests. The principles and detection range are the same for both products.
The following is true for product CS0003:
- Direct procedure for measuring AChE levels in a variety of samples such as serum and plasma.
- Based on the Ellman method in which thiocholine, produced by AChE, reacts with 5,5'-dithiobis (2-nitrobenzoic acid) to form a colorimetric (412 nm) product, proportional to the AChE activity present.
- The AChE activity is measured without the use of a calibration curve. The AChE Positive Control in the kit can be used to screen for AChE inhibitors.
- This kit has a linear range of 10-600 units/liter of AChE activity.
The following is true for product MAK119:
- Direct assays of acetylcholinesterase activity in blood, serum, plasma, and other biological samples.
- 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.
- Detection range 10 to 600 U/L AChE activity in 96-well plate assay.Utile?
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Can I use this AChE kit to test inhibitory substances present in DMSO solution?
1 risposta-
This kit is to measure ACheE activity in blood, serum and plasma samples. It has not been tested to measure AChE levels in a solution of DMSO. If you wanted to test the effects of DMSO present in your sample, you will need to assay a positive control sample alongside a positive control sample spiked with the suspected interfering substance (in this case, DMSO).
Bovine serum can be used as a positive control
Assay buffer can be used as a negative control.Standards can be varying concentrations of acetylcholinesterase within the linear detection range of the kit (10 to 600 U/L AChE activity); one possible source of acetylcholinesterase can be from Sigma Aldrich C3389 (Acetylcholinesterase from electric eel, link: https://www.sigmaaldrich.com/US/en/product/sigma/c3389
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