Invitrogen

Acetylcholinesterase Monoclonal Antibody (HR2)

Product Image Gallery

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

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.

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