Recombinant
RabMAb

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

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 for mouse and rat.

  • Tested applications
    Suitable for: IHC-P, ICC/IF, Flow Cyt, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Choline Acetyltransferase aa 700 to the C-terminus. The exact sequence is proprietary.
    Database link: P28329
    (Peptide available as ab203107)

  • Positive control
    • WB: SH-SY5Y and C6 cell lysate, human fetal brain lysate, mouse and rat brain lysates IP: Human fetal brain lysate FC: SH-SY5Y cells ICC/IF: U-87 MG cells IHC-P: Human cerebrum tissue
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

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

Applications

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.

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.

ICC/IF 1/50 - 1/100.
Flow Cyt 1/60 - 1/110.

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

 

IP 1/30 - 1/70.
WB 1/1000 - 1/10000. Predicted molecular weight: 82 kDa.

Target

Images

  • 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.

  • 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.

  • 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.

  • 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 analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling Choline Acetyltransferase with purified ab181023 at 1/60 dilution (10 µg/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).

  • 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.

  • 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 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.

  • 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.

References

This product has been referenced in:
  • Zhang X  et al. Central cholinergic system mediates working memory deficit induced by anesthesia/surgery in adult mice. Brain Behav 8:e00957 (2018). Read more (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 . Read more (PubMed: 28283678) »
See all 2 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

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

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Verified customer

Submitted Feb 28 2019

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