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  1. Link

    choline-acetyltransferase-antibody-epr13024b-ab181023.pdf

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Neuroscience Neurotransmitter Acetylcholine
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RecombinantRabMAb

Recombinant Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (10)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Immunohistochemistry (Frozen sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
  • Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13024(B)] to Choline Acetyltransferase
  • Suitable for: IHC-P, Flow Cyt (Intra), IHC-Fr, ICC/IF, IP, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free PE

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Overview

  • Product name

    Anti-Choline Acetyltransferase antibody [EPR13024(B)]
    See all Choline Acetyltransferase primary antibodies
  • Description

    Rabbit monoclonal [EPR13024(B)] to Choline Acetyltransferase
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P, Flow Cyt (Intra), IHC-Fr, ICC/IF and IP for mouse and rat.

  • Tested applications

    Suitable for: IHC-P, Flow Cyt (Intra), IHC-Fr, ICC/IF, IP, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: SH-SY5Y and C6 cell lysate, human fetal brain lysate, mouse and rat brain lysates IP: Human fetal brain lysate Flow Cyt (intra): SH-SY5Y cells ICC/IF: U-87 MG and SH-SY5Y cells IHC-P: Human cerebrum tissue IHC-Fr: Hu cerebral cortex tissue sections.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR13024(B)
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Acetylcholine
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab192465)
    • Alexa Fluor® 647 Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab192466)
    • PE Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab224000)
    • APC Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab224001)
    • Anti-Choline Acetyltransferase antibody [EPR13024(B)] - BSA and Azide free (ab224267)
    • Alexa Fluor® 647 Anti-Choline Acetyltransferase antibody [EPR16590] (ab225262)
    • Alexa Fluor® 488 Anti-Choline Acetyltransferase antibody [EPR16590] (ab225263)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181023 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified format use at 1/100 dilution.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Flow Cyt (Intra)
1/60 - 1/110.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr (1)
Use at an assay dependent concentration.
ICC/IF
1/50 - 1/100.
IP
1/30 - 1/70.
WB
1/1000 - 1/10000. Predicted molecular weight: 82 kDa.
Notes
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified format use at 1/100 dilution.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Flow Cyt (Intra)
1/60 - 1/110.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr
Use at an assay dependent concentration.
ICC/IF
1/50 - 1/100.
IP
1/30 - 1/70.
WB
1/1000 - 1/10000. Predicted molecular weight: 82 kDa.

Target

  • Function

    Catalyzes the reversible synthesis of acetylcholine (ACh) from acetyl CoA and choline at cholinergic synapses.
  • Involvement in disease

    Myasthenic syndrome, congenital, 6, presynaptic
  • Sequence similarities

    Belongs to the carnitine/choline acetyltransferase family.
  • Target information above from: UniProt accession P28329 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1103 Human
    • Entrez Gene: 12647 Mouse
    • Entrez Gene: 290567 Rat
    • Omim: 118490 Human
    • SwissProt: P28329 Human
    • SwissProt: Q03059 Mouse
    • SwissProt: P32738 Rat
    • Unigene: 302002 Human
    • Unigene: 442817 Mouse
    • Unigene: 45116 Rat
    see all
  • Alternative names

    • Acetyl CoA choline O acetyltransferase antibody
    • Acetyl CoA:choline O acetyltransferase antibody
    • ChAT antibody
    • CHOACTase antibody
    • Choline acetylase antibody
    • choline acetyltransferase antibody
    • Choline O acetyltransferase antibody
    • Choline O-acetyltransferase antibody
    • CLAT_HUMAN antibody
    • CMS1A antibody
    • CMS1A2 antibody
    • EC 2.3.1.6 antibody
    • OTTHUMP00000019583 antibody
    • OTTHUMP00000019584 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling Choline Acetyltransferase with purified ab181023 at 1/2000 dilution (0.28 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Frozen sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunohistochemistry (Frozen sections) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    IHC image of Choline Acetyltransferase staining in a section of frozen normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab181023, 1/2000 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Immunocytochemistry/ Immunofluorescence analysis of U-87 MG (Human glioblastoma-astrocytoma epithelial cell) cells labeling Choline Acetyltransferase with purified ab181023 at 1/50 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    All lanes : Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023) at 1/2000 dilution (Purified)

    Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates
    Lane 4 : C6 (Rat glial tumor glial cell) whole cell lysates
    Lane 5 : Human fetal brain lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 82 kDa
    Observed band size: 70-82 kDa why is the actual band size different from the predicted?



    This antibody recognizes all Choline Acetyltransferase isoforms with the MW ranging from 70-82 KDa based on immunogen blast.

  • Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Western blot - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    All lanes : Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023) at 1/5000 dilution (unpurified)

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : Human fetal brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 82 kDa



    Blocking/ Dilution buffer: 5% NFDM /TBST.

  • Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Choline Acetyltransferase with purified ab181023 at 1/60 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    ab181023 (purified) at 1/30 dilution (2ug) immunoprecipitating Choline Acetyltransferase in Human fetal brain lysate. Human fetal brain lysate 10ug
    Lane 2 (+): ab181023 & Human fetal brain lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab181023 in Human fetal brain lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunocytochemistry/ Immunofluorescence - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Immunofluorescent analysis of 4% paraformaldehyde fixed SH-SY5Y cells labeling Choline Acetyltransferase using ab181023 (unpurified) at a 1/100 dilution. A Goat anti rabbit IgG (Alexa Fluor®555) was used as the secondary at a 1/100 dilution.

  • Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Flow Cytometry (Intracellular) - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Intracellular Flow Cytometry analysis of SH-SY5Y cells labeling Choline Acetyltransferase using ab181023 (unpurified) at a 1/110 dilution (pink). Goat anti rabbit IgG (FITC) used as the secondary at a 1/150 dilution. Isotype control Rabbit monoclonal IgG (green). Cells were fixed in 2% paraformaldehyde. 

     

     

  • Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Immunoprecipitation - Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

    Lysate from Human fetal brain (Lane 1) and  negative control (Lane 2) were immunoprecipitated with ab181023 (unpurified) at a 1/70 dilution. A anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at a 1/1500 dilution for the secondary. Blocking/ Dilution buffer: 5% NFDM/TBST.

  • Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)
    Anti-Choline Acetyltransferase antibody [EPR13024(B)] (ab181023)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (10)

Publishing research using ab181023? Please let us know so that we can cite the reference in this datasheet.

ab181023 has been referenced in 10 publications.

  • Saw EL  et al. Activation of the cardiac non-neuronal cholinergic system prevents the development of diabetes-associated cardiovascular complications. Cardiovasc Diabetol 20:50 (2021). PubMed: 33618724
  • Ye L  et al. Caspase-11-mediated enteric neuronal pyroptosis underlies Western diet-induced colonic dysmotility. J Clin Invest 130:3621-3636 (2020). PubMed: 32484462
  • Muñoz SS  et al. A Simple Differentiation Protocol for Generation of Induced Pluripotent Stem Cell-Derived Basal Forebrain-Like Cholinergic Neurons for Alzheimer's Disease and Frontotemporal Dementia Disease Modeling. Cells 9:N/A (2020). PubMed: 32887382
  • Jo BG  et al. Vagal afferent fibers contribute to the anti-inflammatory reactions by vagus nerve stimulation in concanavalin A model of hepatitis in rats. Mol Med 26:119 (2020). PubMed: 33272194
  • Catanese A  et al. Retinoic acid worsens ATG10-dependent autophagy impairment in TBK1-mutant hiPSC-derived motoneurons through SQSTM1/p62 accumulation. Autophagy 15:1719-1737 (2019). PubMed: 30939964
  • Jimenez Naranjo C  et al. Effect of cannabidiol on muscarinic neurotransmission in the pre-frontal cortex and hippocampus of the poly I:C rat model of schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry 94:109640 (2019). PubMed: 31108177
  • Chang YX  et al. Intramuscular Injection of Adenoassociated Virus Encoding Human Neurotrophic Factor 3 and Exercise Intervention Contribute to Reduce Spasms after Spinal Cord Injury. Neural Plast 2019:3017678 (2019). PubMed: 30984254
  • Jun H  et al. An immune-beige adipocyte communication via nicotinic acetylcholine receptor signaling. Nat Med 24:814-822 (2018). PubMed: 29785025
  • Zhang X  et al. Central cholinergic system mediates working memory deficit induced by anesthesia/surgery in adult mice. Brain Behav 8:e00957 (2018). PubMed: 29761010
  • Sun P  et al. Deficiency of a7 nicotinic acetylcholine receptor attenuates bleomycin-induced lung fibrosis in mice. Mol Med 23:N/A (2017). Mouse . PubMed: 28283678

Customer reviews and Q&As

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Immunohistochemistry (Frozen sections) abreview for Anti-Choline Acetyltransferase antibody [EPR13024(B)]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Read More

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Submitted Feb 28 2019

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