ACHE Colorimetric Assay Kit

The ACHE Colorimetric Assay Kit is designed to measure the activity of ACHE (acetylcholinesterase) for screening and profiling applications. The ACHE colorimetric reaction is based on a modified Ellman’s method, using the alternative substrate acetylthiocholine iodide (ATCI) and 5,5’-dithio-bis-2-nitrobenzoic acid (DTNB) to quantify amount of thiocholine produced from the hydrolysis of ATCI by ACHE. This assay kit comes in a convenient 96-well format, with purified ACHE enzyme (amino acids 32-614), ATCI, DTNB and assay buffer for 100 enzyme reactions. The absorption intensity of DTNB adduct (λ=410 nm) is used to measure the amount of thiocholine product generated, which is proportional to ACHE activity.Need us to run inhibitor screens or profile your compounds against ACHE? Check out ourMetabolic Enzyme Screening Services.

Product Specifications

Background

ACHE (acetylcholinesterase), also known as AChase or acetylhydrolase, is known to hydrolyze acetylcholine (ACh), a naturally occurring neurotransmitter, into acetic acid and choline. It is a highly effective hydrolase, with an activity near the limit of substrate diffusion. It can be found in motor and sensory neurons, and it is involved in the termination of impulse transmission, by being located on the post-synaptic membrane and hydrolyzing Ach. Ach can be taken up by and used to synthesize acetyl-Coa again in a reaction catalyzed by choline acetyltransferase. ACHE can exist in different molecular forms, which have different expression patterns. It has been known for many years that AD (Alzheimer’s disease) patients have an abnormal distribution of these molecular forms, with an increase in the light forms versus G4 molecules. P-tau can lead to higher expression of T-ACHE, and ACHE may play a role in formation of beta-amyloid plaques. Inhibitors of ACHE result in high concentrations of acetyl-COA and continuous signaling, and if they are irreversible and can lead to muscular paralysis, convulsions, difficulty breathing and asphyxiation. A well-known class of such irreversible inhibitors are organophosphates, that have been used as nerve gases and insecticides. Reversible inhibitors have been approved by the FDA and used to attempt to improve neurological disorders, such as Alzheimer’s disease, myasthenia gravis and Lewy body dementia.

UniProt

P22303

Applications

Screen molecules that inhibit ACHE in drug discovery high-throughput screening (HTS) applications.Determine ACHE inhibitor IC50Perform ACHE activity real-time kinetics analysis

Format

Catalog #NameAmountStorage11004ACHE, His-Tag*>1 µg-80°CDTNB (Lyophilized) 100 reactions-80°CAcetylthiocholine Iodide (ATCI) (Lyophilized) 100 reactions-80°C793113x Immuno Buffer4 ml-80°C79963Transparent 96-well microtiter plate1Room Temp* The initial concentration of enzyme is lot-specific and will be indicated on the tube containing the protein.

Shipping Conditions

-80°C

Storage Conditions

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Notes

Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]