Anti-Ctip2 antibody [25B6] (ab18465)
Key features and details
- Rat monoclonal [25B6] to Ctip2
- Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG2a
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Overview
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Product name
Anti-Ctip2 antibody [25B6]
See all Ctip2 primary antibodies -
Description
Rat monoclonal [25B6] to Ctip2 -
Host species
Rat -
Specificity
Detects 2 bands representing Ctip2 at about 120kD. Ctip2 is highly expressed in brain and in malignant T-cell lines derived from patients with adult T-cell leukemia/lymphoma. -
Tested applications
Suitable for: ICC/IF, WB, Flow Cyt, IHC-Pmore details
Unsuitable for: IHC-Fr -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Recombinant fragment corresponding to Human Ctip2. GTS fusion
Database link: Q9C0K0 -
Epitope
Between amino acids 1-150 of CTIP2. -
Positive control
- Flow Cyt: Jurkat cells. ICC/IF: Neonatal mouse hippocampal cultured neurons, Jurkat cells. WB: Nuclear extract from Jurkat cells; Mouse brain tissue lysate.
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General notes
Hybridoma produced by fusion of a rat lymphocyte and mouse myeloma.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.50
Preservative: 0.02% Sodium azide
Constituents: 0.357% HEPES, 0.87% Sodium chloride -
Concentration information loading...
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Purity
Multi-step Chromatography -
Clonality
Monoclonal -
Clone number
25B6 -
Isotype
IgG2a -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab18465 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 |
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ICC/IF | (10) |
1/500.
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WB | (3) |
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 95 kDa).
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Flow Cyt | (1) |
Use 1µg for 106 cells.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
IHC-P | (10) |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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ICC/IF
1/500. |
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 95 kDa). |
Flow Cyt
Use 1µg for 106 cells. ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Tumor-suppressor protein involved in T-cell lymphomas. May function on the P53-signaling pathway. May be a key regulator of both differentiation and survival during thymocyte development. Repress transcription through direct, TFCOUP2-independent binding to a GC-rich response element. -
Tissue specificity
Highly expressed in brain and in malignant T-cell lines derived from patients with adult T-cell leukemia/lymphoma. -
Sequence similarities
Contains 6 C2H2-type zinc fingers. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 64919 Human
- Entrez Gene: 58208 Mouse
- Omim: 606558 Human
- SwissProt: Q9C0K0 Human
- SwissProt: Q99PV8 Mouse
- Unigene: 709690 Human
- Unigene: 392694 Mouse
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Alternative names
- ATL1 alpha antibody
- ATL1 antibody
- ATL1 beta antibody
see all
Images
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Negative control image: IHC image of Ctip2 staining in a section of formalin-fixed paraffin-embedded mouse cerebellum performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab18465, 5ug/ml, 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. -
IHC image of Ctip2 staining in a section of formalin-fixed paraffin-embedded mouse hippocampus performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab18465, 5ug/ml, 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. -
Negative control image: IHC image of Ctip2 staining in a section of formalin-fixed paraffin-embedded human cerebellum* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab18465, 5ug/ml, 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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
IHC image of Ctip2 staining in a section of formalin-fixed paraffin-embedded human hippocampus* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab18465, 5ug/ml, 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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
Immunofluorescence staining of Ctip2 using ab18465 in Jurkat cells (+ve expression control, top panel) and Daudi cells (-ve expression control, bottom panel). The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton-X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18465 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150084) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunofluorescence staining of Ctip2 using ab18465 in Jurkat cells (+ve expression control, top panel) and Daudi cells (-ve expression control, bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton-X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab18465 at 0.2 µg/mL and ab6046 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150084) (shown in red), both at 1/1000. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Neonatal mouse hippocampal neurons stained with ab18465 (top panel - green) and Ctip1 antibody (middle - red). Bottom panel is overlay of ab18465 and Ctip1 antibody staining - yellow indicates co-localisation, green is ab18465 alone and red is Ctip1 antibody alone.
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Western blot using ab18465 on nuclear extract from Jurkat cells immunoprecipitated with anti-Sir2 antibody.
Western blot using ab18465 on nuclear extract from Jurkat cells immunoprecipitated with anti-Sir2 antibody. Two bands are seen which may correspond to two CTIP2 transcripts present in Jurkat cells as previously reported (Bernard et al. 2001).
Two bands are seen which may correspond to two CTIP2 transcripts present in Jurkat cells as previously reported (Bernard et al. 2001). -
Neonatal Mouse Hippocampal Neurons (Harvested at P1, grown 5d in culture on glial cell feeder layer).
Red is beta tubulin staining.
Green is ab18465. -
Overlay histogram showing Jurkat cells stained with ab18465 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18465, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG (2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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All lanes : Anti-Ctip2 antibody [25B6] (ab18465) at 1/500 dilution
Lanes 1-2 : Mouse brain tissue lysate at 1.5 µg
Lane 3 : Mouse brain tissue lysate at 3 µg
Secondary
All lanes : IRDYE 680-conjugated Donkey Anti-Rat polyclonal. at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 95 kDa
Observed band size: 100,110 kDa why is the actual band size different from the predicted?
Exposure time: 10 minutes
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (718)
ab18465 has been referenced in 718 publications.
- Roşianu F et al. Loss of NDR1/2 kinases impairs endomembrane trafficking and autophagy leading to neurodegeneration. Life Sci Alliance 6:N/A (2023). PubMed: 36446521
- Morelli KH et al. An RNA-targeting CRISPR-Cas13d system alleviates disease-related phenotypes in Huntington's disease models. Nat Neurosci 26:27-38 (2023). PubMed: 36510111
- Junaković A et al. Laminar dynamics of deep projection neurons and mode of subplate formation are hallmarks of histogenetic subdivisions of the human cingulate cortex before onset of arealization. Brain Struct Funct 228:613-633 (2023). PubMed: 36592215
- Liau ES et al. Single-cell transcriptomic analysis reveals diversity within mammalian spinal motor neurons. Nat Commun 14:46 (2023). PubMed: 36596814
- Bandler RC et al. Single-cell delineation of lineage and genetic identity in the mouse brain. Nature 601:404-409 (2022). PubMed: 34912118