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, IHC-Fr, ICC/IF, Flow Cyt, IP, WB
- Reacts with: Mouse, Rat, Human
Overview
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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.
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Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-Fr HumanIHC-P HumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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: SH-SY5Y cells ICC/IF: U-87 MG and SH-SY5Y cells IHC-P: Human cerebrum tissue IHC-Fr: Hu cerebral cortex tissue sections.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13024(B) -
Isotype
IgG -
Research areas
Images
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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.
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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.
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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.
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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.
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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 kDaBlocking/ Dilution buffer: 5% NFDM /TBST.
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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).
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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.
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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.
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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.
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