Butyrylcholinesterase is a nonspecific cholinesterase enzyme that in humans is encoded by the BCHE gene. This gene is mapped to 3q26. It is a member of the type-B carboxylesterase/lipase family of proteins. The encoded enzyme exhibits broad substrate specificity and is involved in the detoxification of poisons including organophosphate nerve agents and pesticides, and the metabolism of drugs including cocaine, heroin and aspirin. Humans homozygous for certain mutations in this gene exhibit prolonged apnea after administration of the muscle relaxant succinylcholine. Sample Type: Cell culture supernates, serum, urine, saliva, human milk, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human BCHE/Butyrylcholinesterase. Sensitivity: <10pg/ml Range: 312pg/ml-20,000pg/ml Specificity: Recognizes natural and recombinant Human BCHE/Butyrylcholinesterase. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human BCHE/Butyrylcholinesterase BioAssay(TM) ELISA Kit was based on standard sandwich enzyme- linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for BCHE/Butyrylcholinesterase has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: E29-L602) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for BCHE/Butyrylcholinesterase is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human BCHE/Butyrylcholinesterase amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human BCHE antibody, 1x plate Lyophilized recombinant Human BCHE standard, 2x10ng/tube Biotinylated anti-Human BCHE antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
