Immunofluorescent analysis of Acetylcholinesterase using Acetylcholinesterase Monoclonal antibody (ZR3) (Product# MA3-041) shows staining in U251 glioma cells. Acetylcholinesterase staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Acetylcholinesterase (Product# MA3-041) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product# 35552 for GAR, Product# 35503 for GAM). Images were taken at 60X magnification.
|Tested species reactivity||Cat, Guinea pig, Human, Mouse, Rabbit, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Purified rat brain acetylcholinesterase.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Blocking Assay (BLOCK)||See 1 publications below|
MA3-041 contains 100 ul of in vitro produced, protein A purified IgG in PBS containing 1 mg/ml BSA and 0.05% sodium azide.
MA3-041 detects acetylcholinesterase (AChE) from feline, human, rat, rabbit and guinea pig tissues. This antibody does not cross react with mouse, chicken, bovine, frog, or electric eel AChE.
MA3-041 has been successfully used in immunohistochemistry and immunoprecipitation procedures. This antibody cannot be used in Western blot to detect AChE. Immunohistochemical staining of Ach in rat brain with MA3-041 results in staining of nerve fibers and terminals. MA3-041 reacts with both the soluble and membrane-associated AChE.
The MA3-041 antigen is purified rat brain acetylcholinesterase.
Cholinergic neurotransmission occurs in motor, autonomic and central nervous synapses and requires very rapid inactivation of its transmitter, acetylcholine (ACh). Acetylcholinesterase (AChE) rapidly hydrolyzes ACh to acetate and choline, thereby inactivating it. AChE is found in the neuromuscular junction anchored to the basal lamina which runs between the nerve terminal and muscle membrane. AChE is also found outside the nervous and neuromuscular system in blood, lymph, germ and liver cells suggesting a role for AChE not related to cholinergic transmission. Another less specific cholinesterase, butyrylcholinesterase (BChE), seems to contribute to the regulation of the ACh concentration in the synaptic cleft.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Paraoxon toxicity is not potentiated by prior reduction in blood acetylcholinesterase.
MA3-041 was used in blocking/activating experiment to investigate the influence of blood acetylcholinesterase reduction on paraoxon toxicity
|Padilla S,Moser VC,Pope CN,Brimijoin WS||Toxicology and applied pharmacology (117:110)||1992|