Description :
Cholinesterase (ChE) consists of a group of enzymes that hydrolyze choline esters. There are two ChE isoenzymes in blood: acetylcholinesterase (AChE; EC 3.1.1.7), also known as erythrocytes or true ChE, which is found mainly in red blood cells; and butyrylcholinesterase (BChE; EC 3.1.1.8), also known as plasma ChE or pseudo-ChE, which is present in plasma. Blood AChE or BChE activity would be selectively reduced by exposing them to poisonous chemical agents, insecticides such as organophosphates or carbamates, anesthetics, and a variety of therapeutic drugs including donepezil or rivastigmine which are used for treating Alzheimer’s diseases. Therefore, Blood Cholinesterases (ChE=AChE+BChE) are potential biomarkers of suppressed and/or increased central and peripheral nervous system activity and tools for confirming possible therapeutics. Since plasma BChE and erythrocyte AChE can be selectively inhibited by certain insecticides or drugs, quantification of both isoenzymes' activities is important. cholinesterase activity kit combines the specific AChE and BChE substrates and a selective BChE inhibitor to measure and distinguish AChE and BChE activities in Whole Blood samples without separating plasma from erythrocytes. The principle is based on the ability of AChE and BChE to hydrolyze their respective substrates and produce thiocholine. Thiocholine reacts with 5,5’-dithiobis(2-nitrobenzoic acid) (DTNB) generating a yellow chromophore (TNB) that can be quantified at 412 nm. It is simple, easy to implement, and useful in clinical research to monitor exposure to anti-ChE compounds in Blood Samples.
Applications :
Measurement of ChE activity in various tissues/cells
Screening of ChE inhibitors
Storage :
Store Kit at -20°C, protect from light. Briefly centrifuge small vials prior to opening. Read entire protocol before performing the assay.
ChE Assay Buffer: Store at 4 ºC or -20 ºC. Bring to room temperature before use.
AChE Substrate: Reconstitute in 100 µl ChE Assay Buffer. Store at -20ºC. Use within two months.
BChE Substrate: Store at -20°C, protect from light. Bring to room temperature before use.
Acetylcholinesterase: Reconstitute with 100 µl of ChE Assay Buffer. Aliquot and store at -20 ºC. Use within two months.
Butyrylcholinesterase: Reconstitute with 20 µl ChE Assay Buffer. Store at -20 ºC. Use within two months.
BChE Inhibitor: Reconstitute BChE Inhibitor in 150 µl dH2O. Vortex intensively at room temperature to facilitate solubilization. Aliquot and store at -20ºC. Bring to room temperature before use. Use it within two months.
DTNB Solution: Dissolve 1 vial of DTNB with 625 µl ChE Assay Buffer. Each vial can be used to carry out up to 50 reactions. Dissolve
vial contents when needed. Store at -20ºC. Use within two months.
TNB Standard: Dissolve in 1 ml of ChE Assay Buffer to generate 2.5 mM TNB Standard. Use within two months.
Kit Components :
AChE Assay Buffer;
AChE Substrate;
BChE Probe (in DMSO);
Acetylcholinesterase;
Butyrylcholinesterase;
BChE Inhibitor;
DTNB;
TNB Standard
Target Species :
Mammalian
Detection method :
Absorbance (OD 412 nm)
Compatible Sample Types :
Biological Fluids: Blood
Features & Benefits :
Rapid, simple & convenient;
This assay kit can detect cholinesterase activity as low as 0.5 mU/ml in a variety of samples
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