Anti Acetylcholinesterase (human brain, bovine brain, AChE)
mouse monoclonal antibody
Cat.No. HYB 190-01
Preparation: Protein-A/G purified
Content: Available in 200 µL and 1 mL
Concentration: 1 mg/mL
Solvent: 0.01 M phosphate buffer, pH 7.4, with 0.5 M NaCl and 15mM sodium azide
Storage: In the dark at 4-8ºC
Acetylcholinesterase (AChE, EC.3.1.1.7.) is an enzyme located in the postsynaptic membrane and in the muscle endplates, where it hydrolyses the neurotransmitter acetylcholin. AChE from brain is a tetramer (G4-AChE) with a molecular mass of 320 kDa, AChE from erythrocytes is a dimer (G2-AChE) with a molecular mass of 170 kDa.
Detection of higher levels of AChE in amniotic fluid can indicate fetal malformations such as neural tube defects.
C-terminal 10 residues of brain acetylcholinesterase (human and bovine), absent from the erythrocyte enzyme.
HYB 190-01 is specific for brain AChE and does not recognize AChE from erythrocytes. The antibody can thus distinguish between mammalian brain AChE and erythrocyte AChE. Weak cross-reactivity with Torpedo marmorata AChE but none with AChE from electric eel or human BtChE.
C-terminal 10 residues (aa 574-583) of brain acetylcholinesterase (DS-AChE and SS-AChE) (1).
Can be used in ELISA on amniotic fluid for the diagnosis of neural tube defects. HYB 190-01 is well suited as catching antibody (on an anti-mouse IgG coat) in enzyme antigen immunoassay (EAIA), where the antigen (AChE) is captured and used directly as substrate for acetylthiocholiniodide (Ellmann’s reaction) (1,2).
In Western blotting and dot blotting HYB 190-01 reacts with native and denatured human and bovine, detergent soluble and salt-soluble AChE. No cross-reactivity is seen with erythrocyte AChE.
| Method | Usability | Dilution guideline | References |
| ELISA | Yes | 1/16,000 | 1 |
| Immunoblotting | Yes | 1/75 | 1 |
| Immunohistochemistry | Not determined |
The dilution guideline for ELISA is based on plates coated with anti-mouse antibody to catch the monoclonal antibody followed by recomb. AChE (human), 20 U/ml. Detection is done with Ellmans reagent. Users should determine the optimal dilution for their own pupose.
1. Boschetti N, Brodbeck U, Jensen SP, Koch C, Norgaard-Pedersen B (1996) Monoclonal antibodies against a C-terminal peptide of human brain acetylcholinesterase distinguish between erythrocyte and brain acetylcholinestrases. Clin Chem 42:19-23.
2. Rasmussen AG, Arends J, Larsen SO (1989) Evaluation and quality control of a monoclonal antibody based enzyme antigen immunoassay of acetylcholinesterase in amniotic fluid. Scand J Clin Lab Invest 49:503-11.
3. Aziz-Aloya R, Sternfeld M, Soreq H (1993) Promoter elements and alternative splicing in the human ACHE gene. Prog Brain Res 98:147-153.
4. Sorensen K, Gentinetta R, Brodbeck U (1982) An amphiphile-dependent form of human brain caudate nucleus acetylcholinesterase: purification and properties. J Neurochem 39:1050-1060.
