Invitrogen

Acetylcholinesterase Monoclonal Antibody (HR2)

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Acetylcholinesterase Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of Acetylcholinesterase using Anti-Acetylcholinesterase Monoclonal Antibody (HR2) (Product # MA3-042) shows staining in Neuro-2a 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 # MA3-042) at a dilution of 1:200 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Acetylcholinesterase Antibody in Immunohistochemistry (IHC) — Immunohistochemistry was performed on normal deparaffinized human Cerebellum tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Acetylcholinesterase (Product # MA3-042) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Acetylcholinesterase Antibody in Immunocytochemistry, Immunohistochemistry (ICC/IF, IHC) — Fig 3 iPSC-derived neurons repress HIV expression. ( A ) 60,000 hmuglia/HIV HC69 cells were plated in the presence of 0.5 x 10 6 LUHMES-derived neurons (as positive control) or 0.5 x 10 6 iPSC-derived GABAergic cortical (iCort), dopaminergic (iDopaNer) or motor neurons (iMotorNer). HIV expression was evaluated after 24 h by fluorescence microscopy. Microglia identified by phase contrast microscopy are outlined by the white contours. ( B ) Flow cytometric analysis of microglial cell GFP expression. The p -values of pair-sample, Student's t -tests comparing the microglial cells cultured alone or in the presence of neurons are shown. Individual independent experiments are color coded ( n = number of independent samples). N.S.: non- s ignificant. ( C ) Identification of differentiation of iPSC-derived neurons. Super-imposed images of DAPI stained nuclei (blue) and Alexa-Fluor 488 stained neuronal antigens (green) are shown. Neurons were stained with antibodies against GAD65/67, DAT, and AchE and Alexa Fluor 488-conjugated anti-rabbit secondary antibody.

Acetylcholinesterase Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 1 Plasma AChE levels in healthy controls (wild-type) after BuChE immunodepletion and in BuChE-silent individuals. ( A ) Control plasma was immunoprecipitated with anti-BuChE antibody and cholinesterase activity levels determined before b and after a immunoprecipitation (n = 6; 46+-4 yrs). AChE activity level in plasma from BuChE-silent individuals is also shown (n = 3; 30+-5 yrs). The anti-BuChE antibody does not immunoprecipitate AChE in BuChE-silent plasma (not shown). Values are means +- SEM. ( B ) Immunoprecipitation of control plasma with antibody, followed by immunoblotting with the anti-AChE antibody, N-19. The presence (+) or absence (-) of the anti-BuChE antibody linked to the resin is indicated in the top margin. Prior to electrophoretic analysis, proteins abundant in plasma were depleted by immunoaffinity-based protein subtraction chromatography with IgY microbeads (Seppro(tm)). The anti-BuChE antibody does not immunoprecipitate AChE. Extracts incubated with protein A-Sepharose, in the absence of the antibody, were analyzed in parallel as negative controls. ( C ) Representative profiles of AChE and ( D ) BuChE molecular forms (G 4 = tetramers; G 1 +G 2 = monomers and dimers) in control plasma samples before (*) and after (*) BuChE-immunoprecipitation, and in BuChE-silent plasma (^). ( E ) Representative immunoblot of individual AChE G 4 and G 1 +G 2 peak-fractions separated by sucrose gradient centrifugation from control plasma and detected with the N-19 ant

Acetylcholinesterase Antibody in Western Blot (WB) — Figure 5 Altered AChE immunoreactivity in plasma of non-demented controls (ND) and Alzheimer's disease (AD) patients. ( A ) Representative blot of plasma-AChE from AD and ND using the antibody N-19, and ( B ) densitometric quantification of the AChE-immunoreactive bands, expressed in arbitrary units (a u.), from 15 ND (#; n = 15) and 14 AD (#; n = 14) subjects. Proteins abundant in plasma were depleted by immunoaffinity-based protein subtraction chromatography with IgY microbeads (Seppro(tm)) and equivalent amounts of protein were loaded in each lane. Columns represent means +- SEM * p = 0.02 significantly different from NDs as assessed by Student's t test. ( C ) Representative blot of plasma AChE detected with the anti-AChE antibody to the C-terminal, ab31276.

Product Details

MA3-042

Applications	Tested Dilution	Publications
Western Blot (WB)	-	View 4 publications 4 publications
Immunohistochemistry (IHC)	-	View 8 publications 8 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:20-1:200	-
Immunohistochemistry (Frozen) (IHC (F))	1:50	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 2 publications 2 publications
ELISA (ELISA)	Assay-dependent	View 1 publication 1 publication
Immunoprecipitation (IP)	Assay-dependent	View 1 publication 1 publication

Product Specifications
Species Reactivity	Bovine, Cat, Guinea pig, Human, Mouse, Rabbit
Published species	Guinea pig, Human, Mouse, Non-human primate
Host/Isotype	Mouse / IgG2b
Class	Monoclonal
Type	Antibody
Clone	HR2
Immunogen	Purified human cerebellar acetylcholinesterase.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_325478

Product Specific Information

MA3-042 detects acetylcholinesterase (AChE) from human, mouse, rabbit, guinea pig, bovine and cat tissues. This antibody does not cross react with rat or frog AChE, and does not detect butyrylcholinesterase (BChE).

MA3-042 has been successfully used in immunohistochemistry, immunoprecipitation and ELISA procedures. This antibody cannot be used in Western blot to detect AChE. Immunohistochemical staining of AChE in human brain samples with MA3-042 results in staining of nerve fibers and terminals.

The MA3-042 antigen is purified human cerebellar acetylcholinesterase.

Target Information

Acetylcholinesterase hydrolyzes the neurotransmitter, acetylcholine at neuromuscular junctions and brain cholinergic synapses, and thus terminates signal transmission. The Protein is also found on the red blood cell membranes, where it constitutes the Yt blood group antigen. Acetylcholinesterase exists in multiple molecular forms which possess similar catalytic properties, but differ in their oligomeric assembly and mode of cell attachment to the cell surface. The major form of acetylcholinesterase found in brain, muscle and other tissues is the hydrophilic species, which forms disulfide-linked oligomers with collagenous, or lipid-containing structural subunits.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Cross-talk between microglia and neurons regulates HIV latency.

PLoS pathogens

Alvarez-Carbonell D,Ye F,Ramanath N,Garcia-Mesa Y,Knapp PE,Hauser KF,Karn J

Published figure using Acetylcholinesterase monoclonal antibody (Product # MA3-042) in Immunofluorescence

Sun Dec 01 00:00:00 EST 2019

Cross-talk between microglia and neurons regulates HIV latency.

PLoS pathogens

Alvarez-Carbonell D,Ye F,Ramanath N,Garcia-Mesa Y,Knapp PE,Hauser KF,Karn J

Published figure using Acetylcholinesterase monoclonal antibody (Product # MA3-042) in Immunofluorescence

Sun Dec 01 00:00:00 EST 2019

Adventitial Cell Atlas of wt (Wild Type) and ApoE (Apolipoprotein E)-Deficient Mice Defined by Single-Cell RNA Sequencing.

Arteriosclerosis, thrombosis, and vascular biology

Gu W,Ni Z,Tan YQ,Deng J,Zhang SJ,Lv ZC,Wang XJ,Chen T,Zhang Z,Hu Y,Jing ZC,Xu Q

MA3-042 was used in Immunohistochemistry-immunofluorescence to study the role of the cellular atlas of the heterogeneous adventitial cells, and the intracellular communication, in atherosclerosis.

Sat Jun 01 00:00:00 EDT 2019

Identification of Carboxylesterase, Butyrylcholinesterase, Acetylcholinesterase, Paraoxonase, and Albumin Pseudoesterase in Guinea Pig Plasma through Nondenaturing Gel Electrophoresis.

Comparative medicine

Napon G,Dafferner AJ,Saxena A,Lockridge O

MA3-042 was used in Immunohistochemistry to identify the major esterases in guinea pig plasma.

Mon Oct 01 00:00:00 EDT 2018

Immunopurification of Acetylcholinesterase from Red Blood Cells for Detection of Nerve Agent Exposure.

Chemical research in toxicology

Dafferner AJ,Schopfer LM,Xiao G,Cashman JR,Yerramalla U,Johnson RC,Blake TA,Lockridge O

MA3-042 was used in Western Blotting to develop a method of immunopurifying human red blood cell acetylcholinesterase in quantities adequate for detecting exposure to nerve agents and organophosphorus pesticides, by mass spectrometry.

Mon Oct 16 00:00:00 EDT 2017

Functional variability in butyrylcholinesterase activity regulates intrathecal cytokine and astroglial biomarker profiles in patients with Alzheimer's disease.

Neurobiology of aging

Darreh-Shori T,Vijayaraghavan S,Aeinehband S,Piehl F,Lindblom RP,Nilsson B,Ekdahl KN,Långström B,Almkvist O,Nordberg A

MA3-042 was used in ELISA to study the effect of Alzheimer's disease patient butyrylcholinesterase genotype on the profile of inflammatory cytokines and markers of astroglial function in CSF

Fri Nov 01 00:00:00 EDT 2013

Lesions of cortical GABAergic interneurons and acetylcholine neurons in xeroderma pigmentosum group A.

Brain & development

Hayashi M,Ohto T,Shioda K,Fukatsu R

MA3-042 was used in immunohistochemistry to study the status of cortical GABAergic interneurons and cholinergic neurons in xeroderma pigmentosum group A

Sun Apr 01 00:00:00 EDT 2012

Decrease in acetylcholinergic neurons in the pedunculopontine tegmental nucleus in a patient with Prader-Willi syndrome.

Neuropathology : official journal of the Japanese Society of Neuropathology

Hayashi M,Miyata R,Tanuma N

MA3-042 was used in immunohistochemistry to study the selective deficiency of acetylcholine neurons in the pedunculopontine tegmental nucleus with Prader-Willi syndrome

Wed Jun 01 00:00:00 EDT 2011

Altered levels of acetylcholinesterase in Alzheimer plasma.

PloS one

García-Ayllón MS,Riba-Llena I,Serra-Basante C,Alom J,Boopathy R,Sáez-Valero J

MA3-042 was used in immunoprecipitation to investigate the changes of acetylcholinesterase level in Alzheimer plasma

Thu Jan 14 00:00:00 EST 2010

Altered levels of acetylcholinesterase in Alzheimer plasma.

PloS one

García-Ayllón MS,Riba-Llena I,Serra-Basante C,Alom J,Boopathy R,Sáez-Valero J

MA3-042 was used in immunoprecipitation to investigate the changes of acetylcholinesterase level in Alzheimer plasma

Thu Jan 14 00:00:00 EST 2010

Different cholinesterase inhibitor effects on CSF cholinesterases in Alzheimer patients.

Current Alzheimer research

Nordberg A,Darreh-Shori T,Peskind E,Soininen H,Mousavi M,Eagle G,Lane R

MA3-042 was used in western blot to investigate the effect of cholinesterase inhibitors on the actvities and protein levels of cholinesterases

Sun Feb 01 00:00:00 EST 2009

Presenilin 1 interacts with acetylcholinesterase and alters its enzymatic activity and glycosylation.

Molecular and cellular biology

Silveyra MX,Evin G,Montenegro MF,Vidal CJ,Martínez S,Culvenor JG,Sáez-Valero J

MA3-042 was used in immunocytochemistry, immunoprecipitation and western blot to indicate that PS1 can interact with AChE and influence its expression

Thu May 01 00:00:00 EDT 2008

Presenilin 1 interacts with acetylcholinesterase and alters its enzymatic activity and glycosylation.

Molecular and cellular biology

Silveyra MX,Evin G,Montenegro MF,Vidal CJ,Martínez S,Culvenor JG,Sáez-Valero J

MA3-042 was used in immunocytochemistry, immunoprecipitation and western blot to indicate that PS1 can interact with AChE and influence its expression

Thu May 01 00:00:00 EDT 2008

Presenilin 1 interacts with acetylcholinesterase and alters its enzymatic activity and glycosylation.

Molecular and cellular biology

Silveyra MX,Evin G,Montenegro MF,Vidal CJ,Martínez S,Culvenor JG,Sáez-Valero J

MA3-042 was used in immunocytochemistry, immunoprecipitation and western blot to indicate that PS1 can interact with AChE and influence its expression

Thu May 01 00:00:00 EDT 2008

Fetal fornix transection and gestation length in sheep.

Experimental neurology

McDonald TJ,Li C,Vincent SE,Nijland MJ

MA3-042 was used in immunohistochemistry to study the developmental state of fetal hippocampus and its role in gestation

Tue Aug 01 00:00:00 EDT 2006

Immunocytochemical demonstration of axonal and perikaryal acetylcholinesterase in human cerebral cortex.

Brain research

Mesulam MM,Geula C,Cosgrove R,Mash D,Brimijoin S

MA3-042 was used in immunohistochemistry to investigate the expression of acetylcholinesterase in axons and perikarya in the human cerebral cortex

Fri Jan 25 00:00:00 EST 1991

Bioinformatics

Protein Aliases: Acetylcholinesterase; acetylcholinesterase (Yt blood group); acetylcholinesterase T-subunit; AChE; apoptosis-related acetylcholinesterase; EC 3.1.1.7; OTTHUMP00000211349; OTTHUMP00000211356; Yt blood group

Gene Aliases: ACEE; ACHE; ARACHE; N-ACHE; YT

UniProt ID: (Human) P22303, (Bovine) P23795, (Cat) O62763, (Mouse) P21836

Entrez Gene ID: (Rabbit) 100009390, (Human) 43, (Bovine) 540446, (Cat) 493674, (Mouse) 11423, (Guinea pig) 100422792

Molecular Function: esterase