
- Description
- Additional Information
- Readable Documents
- Assay Principle
- Reviews
Introduction
Exposure to chemical nerve agents, pesticides and certain drugs (anesthetics, cocaine and therapeutical drugs) reduces the activity of red blood cell (RBC) acetylcholinesterase (AChE). The RBC-AChE can be used as a biomarker to monitor suppressed and or increased AChE function in the peripheral and central nervous system (9). Acetylcholinesterase (AChE) is one of the most important enzymes involved in nerve transmission. The enzyme is bound to cellular membranes of excitable tissue (synaptic junction, endoplasmic reticulum, etc) 1-3. Acute toxicity to humans and animals through inhibition of AChE by both nerve gases and an important class of pesticides has long been a field of intensive scientific investigation 4,5. AChE inhibitors have also been used clinically as Alzheimer’s treatments (e.g., tacrine (tetrahydroaminoacridine)) 6 and are the subject of increasing interest in various disease processes and treatment strategies 7,8. However, both environmental detection of AChE inhibitors and development of modulators of AChE enzymatic activity as drugs have been hampered by the difficulty and complexity of the current assay methods.
Key Benefits
- Non Radioactive assay can monitor multiple time points to follow kinetics.
- One-step, no wash assay.
- Versatile: Can Detect AChE activity in RBC’s, Saliva and Tissue Lysates.
- Readout – 96 well Fluorescent Plate Reader.
Additional information
Kit Size | 100, 500 |
---|
We have developed a highly sensitive, very rapid, extremely simple assay to determine acetylcholinesterase activity in RBC’s, using the natural substrate, acetylcholine. Additionally, by using specific inhibitors, the kit can be used to detect AChE activity in a variety of samples. A series of coupled enzyme reactions quickly translates the presence of active AChE into a change in the fluorescence of a quenched detection reagent.
AChE + ATP + H20 + coupled enzyme reaction + quenched dye —-> Fluorescent
Dye (Ex:530-570nm
Em:590-600nm
Time=10 min | Dilution | Volume used | ug protein/well | RFU * |
---|---|---|---|---|
RBC | 1:1000 | 10uL | 5 | 3450 |
RBC no Ach | 1:1000 | 10uL | 5 | 152 |
Rat brain | 1:500 | 10uL | 20 | 5864 |
Rat brain no Ach | 1:500 | 10uL | 20 | 296 |
Saliva | neat | 10uL | ND | 925 |
Saliva no Ach | neat | 10uL | ND | 772 |
RBC-AChE mU/mL in well | RFU | SD RFU |
---|---|---|
167.000 | 9999 | 0.00 |
41.750 | 6375 | 121.70 |
10.438 | 1659 | 9.00 |
2.609 | 590 | 9.81 |
0.652 | 256 | 7.23 |
0.163 | 222 | 8.72 |
0 | 186 | 8.08 |
Document Title |
Fluoro AChE Protocol |
Fluoro AChE Datasheet |
msds.fluoroAChE |
Title | File | Link | Author(s) | Journal | Year; Edition:Pages |
Gonadal hormones modulate the potency of the disruptive effects of donepezil in male rats responding under a nonspatial operant learning and performance task | http://journals.lww.com/behaviouralpharm/Abstract/publishahead/Gonadal_hormones_modulate_the_potency_of_the.99913.aspx | Leonard, Stuart T.; Hearn, John K.; Catling, Andrew D.; Winsauer, Peter J | Behavioral Pharmacology | Feb 2010 | |
Acetylcholinesterase, a key prognostic predictor for hepatocellular carcinoma, suppresses cell growth and induces chemosensitization | http://onlinelibrary.wiley.com/doi/10.1002/hep.24079/full | Yingjun Zhao, Xiaoying Wang, Tao Wang, Xin Hu, Xin Hui, Mingxia Yan, Qiang Gao, Taoyang Chen, Jinjun Li, Ming Yao, Dafang Wan, Jianren Gu, Jia Fan, Xianghuo He | Hepatology | Vol 53, Issue 2, pp 493–503, February 2011-DOI: 10.1002/hep.24079 |
Reference |
Politoff, A., Blitz, A., and Rose, S.: Incorporation of Acetylcholinesterase Into Synaptic Vesicles is Associated with Blockade of Synaptic Transmission, Nature 256, 324, 1975 |
Friedenberg, R., and Seligman, A.: Acetylcholinesterase at the Myoneural Junction: Cytochemical Ultrastructure and Some Biochemical Considerations, J Histochem Cytochem 20, 771, 1972 |
Nachmansohn, D.: Proteins in Excitable Membranes, Science 168, 1059, 1970. |
HA Berman and MM Decker Kinetic, equilibrium, and spectroscopic studies on dealkylation ("aging") of alkyl organophosphonylacetylcholinesterase. Electrostatic control of enzyme topography. J. Biol. Chem., Aug 1986; 261: 10646-10652 . |
Arie Ordentlich et al. The Architecture of Human Acetylcholinesterase Active Center Probed by Interactions with Selected Organophosphate Inhibitors. J. Biol. Chem., May 1996; 271: 11953-11962. |
Levy R. Tetrahydroaminoacridine and Alzheimer"s disease. Lancet, 1987 Feb 7;1(8528):322. |
Bolognesi ML et al. Propidium-based polyamine ligands as potent inhibitors of acetylcholinesterase and acetylcholinesterase-induced amyloid-beta aggregation. J Med Chem. 2005 Jan 13;48(1):24-7. |
Schallreuter KU et al. Activation/deactivation of acetylcholinesterase by H2O2: more evidence for oxidative stress in vitiligo. Biochem Biophys Res Commun. 2004 Mar 5;315(2):502-8. |
# Nigg HN, Knaak JB. Blood cholinesterases as human biomarkers of organophosphorus pesticide exposure. Rev. Environ. Contam. Toxicol, 2000:163: p29-111. |
Part# | Reagent | Temperature |
Part# 3023 | Component A: Detection Reagent Diluent, 5.5mL | -20C |
Part# 3024 | Component B: Coupled Enzyme Reagent, 5.5mL | -20C |
Part# 4016 | Component C: Detection Reagent, 1 Vial | -20C |
Part# 3011 | Component D: 5X Reaction Buffer | -20C |
Part# 7012 | Component E: Acetylcholine | -20C |
Part# 6020 | Component E: Reg Blood Cell Acetylcholinesterase | -20C |
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⒈通过嵌入DNA双链的碱基之间,形成稳定复合物,抑制DNA复制与RNA合成,从而阻碍快速生长的癌细胞的分裂。
⒉抑制拓扑异构酶II,影响DNA超螺旋转化成为松弛状态,从而阻碍DNA复制与转录。有研究显示拓扑异构酶II抑制剂(除蒽环类药物还包括依托泊苷等)能够阻止拓扑异构酶II的翻转,而这点对于它从它的核酸底物上脱离是必需的。这就意味着,拓扑异构酶II抑制剂使拓扑异构酶II的复合物在DNA链断裂之后才能更稳定,导致后者催化了DNA的破坏;同时,拓扑异构酶II抑制剂还能阻碍连接酶对DNA的修复。
⒊螯合铁离子后产生自由基从而破坏DNA、蛋白质及细胞膜结构。向左转|向右转
ResponseofskeletalmuscleRNApolymerasesIandIItotumourgrowth
BiochimicaetBiophysicaActa(BBA)-GeneStructureandExpression,Volume950,Issue3,7September1988,Pages296-302
GeorgeA.J.Goodlad,CatherineM.Clark
或者有高手知道怎么测也行
最近遇到了实验上的大问题,就是用一步法做RNA的荧光定量PCR时,发现低浓度的做不出来,而DNA是可以的,想请问各位在一步法PCR中有没有什么高见,就是逆转录的Buffer有没有什么需要注意的地方!就是在逆转录酶的最适Buffer和Taq酶的最适Buffer中寻找一个最适的Buffer!
我是新手。我要开始做实验了,但一开始就碰到了个很大的问题。
我想买个能利用哺乳动物细胞RNA聚合酶将DNA转录成RNA的的质粒,向好几家公司打了电话,但都没有我想要的东西。
没有这个东西,我的实验就只能是蓝图了。哪位战友有办法的,能不能帮帮我?
不胜感激。
请规范发贴,求助贴请在标题前加【求助】,谢谢您的合作。——by草根

