Acetylcholinesterase Reagent

Product Name:	Acetylcholinesterase Reagent (BA0208)
SKU:	BA0208
Detection Method:	Colorimetric (OD 410 nm; improved Ellman method)
Detection Range:	6 - 750 U/L
Sample Type:	Whole blood, serum, EDTA plasma and other biological samples
Species Reactivity:	All
Assay Time:	10 minutes (kinetic read at 1 min and 10 min)
Kit Size:	100 Assays
Equipment Required:	Microplate reader
Storage:	Shipped at room temperature. Store unopened at 2-25 degrees C in a dry environment.
Shelf Life:	6 months after receipt
Shipping:	Room Temperature

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 catalyses 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 organic phosphate insecticides. This reagent is based on an improved Ellman method, in which thiocholine produced by the action of acetylcholinesterase forms a yellow colour with 5,5'-dithiobis(2-nitrobenzoic acid). The intensity of the product colour at 410 nm is proportional to the enzyme activity in the sample.

Simple and convenient single reagent that is ready to use once reconstituted in 25 mL of deionised water.
High-throughput. Can be readily adapted on chemistry analysers or high-throughput liquid handling systems to test thousands of samples per day.
Detection range 6 - 750 U/L.

Direct assays of acetylcholinesterase activity in blood, serum, plasma and other biological samples.

Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Step	Procedure
1	Before using the reagent, lightly tap it to settle the powder, add 25 mL of distilled or deionised water, cap the bottle and thoroughly mix to dissolve the contents by shaking for 10 seconds; the reconstituted Working Reagent is a pale yellowish liquid, pH 7.2.
2	Prepared reagent is stable for 8 hours at room temperature or 3 days at 2-8 degrees C, protected from light.
3	Transfer 5 uL of calibrator into separate wells of a clear-bottom 96-well plate.
4	Transfer 5 uL of each sample per well into separate wells.
5	Transfer 225 uL of freshly prepared Working Reagent to all assay wells and tap the plate briefly to mix.
6	Read OD410nm at 1 minute and at 10 minutes in a plate reader, or alternatively record kinetics at 410 nm.

Acetylcholinesterase activity is calculated as: ACHE Activity = [(OD₁₀ - OD₁)_SAMPLE / (OD₁₀ - OD₁)_CAL] x Cal (U/L), where OD₁₀ and OD₁ are the OD_410nm values at 10 minutes and 1 minute respectively, and Cal (U/L) is the AChE activity of the calibrator used. If the calculated AChE activity is higher than 750 U/L, dilute the sample in 0.9% NaCl, repeat the assay and multiply the result by the dilution factor. Unit definition: one unit of enzyme catalyses the production of 1 umole of thiocholine per minute under the assay conditions (pH 7.2 and room temperature).

Component	Quantity	Storage
Reagent (powder in one 30 mL amber bottle)	sufficient for approximately 100 tests	2-25 degrees C, dry

Acetylcholinesterase Reagent (BA0208)

Description