Recombinant Anti-ATF1 antibody [EPR17028] (ab181569)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17028] to ATF1
- Suitable for: IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ATF1 antibody [EPR17028]
See all ATF1 primary antibodies -
Description
Rabbit monoclonal [EPR17028] to ATF1 -
Host species
Rabbit -
Tested applications
Suitable for: IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse heart and kidney lysates; Rat heart lysate; HeLa, C6, RAW 264.7 and NIH/3T3 whole cell lysates. IHC-P: Mouse liver tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells. IP: Mouse kidney lysate.
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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.
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: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17028 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181569 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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IP |
1/50.
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WB |
1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 29 kDa).
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IHC-P |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This antibody is not suitable for human and rat species in IHC application. |
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ICC/IF |
1/100.
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Flow Cyt (Intra) |
1/150.
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Notes |
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IP
1/50. |
WB
1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 29 kDa). |
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This antibody is not suitable for human and rat species in IHC application. |
ICC/IF
1/100. |
Flow Cyt (Intra)
1/150. |
Target
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Function
This protein binds the cAMP response element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Binds to the Tax-responsive element (TRE) of HTLV-I. Mediates PKA-induced stimulation of CRE-reporter genes. -
Involvement in disease
Defects in ATF1 may be a cause of angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. A distinct variant of malignant fibrous histiocytoma that typically occurs in children and adolescents and is manifest by nodular subcutaneous growth. Characteristic microscopic features include lobulated sheets of histiocyte-like cells intimately associated with areas of hemorrhage and cystic pseudovascular spaces, as well as a striking cuffing of inflammatory cells, mimicking a lymph node metastasis. Note=Chromosomal aberrations involving ATF1 are found in patients with angiomatoid fibrous histiocytoma. Translocation t(12;16)(q13;p11.2) with FUS generates a chimeric ATF1/FUS protein. Translocation t(12;22)(q13;q12) with EWSR1 generates a chimeric ATF1/EWSR1 protein. -
Sequence similarities
Belongs to the bZIP family. ATF subfamily.
Contains 1 bZIP domain.
Contains 1 KID (kinase-inducible) domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 466 Human
- Entrez Gene: 11908 Mouse
- Entrez Gene: 315305 Rat
- Omim: 123803 Human
- SwissProt: P18846 Human
- SwissProt: P81269 Mouse
- Unigene: 648565 Human
- Unigene: 473546 Mouse
see all -
Alternative names
- Activating transcription factor 1 antibody
- ATF 1 antibody
- atf1 antibody
see all
Images
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All lanes : Anti-ATF1 antibody [EPR17028] (ab181569) at 1/1000 dilution
Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat heart lysate
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:10574952).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling ATF1 with ab181569 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling ATF1 with ab181569 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on mouse liver (PMID: 11865068; PMID: 28032861). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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ATF1 was immunoprecipitated from 1 mg of mouse kidney lysate with ab181569 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab181569 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: Mouse kidney lysate 10 µg (Input).
Lane 2: ab181569 IP in mouse kidney lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181569 in mouse kidney lysate (-).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling ATF1 with ab181569 at 1/150 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-ATF1 antibody [EPR17028] (ab181569) at 1/1000 dilution
Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 3 minutes; Lane 2: 30 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-ATF1 antibody [EPR17028] (ab181569) at 1/1000 dilution
Lane 1 : Mouse kidney lysate
Lane 2 : Mouse kidney lysate treated with alkaline phosphatase for 1 hour at 37°C
Lysates/proteins at 2.5 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
ATF1 has been shown to be phosphorylated in cultured cells and hyperphosphorylation in tissues (PMID:20730097). Treatment of cell lysates with alkaline phosphatase lead to complete dephosphorylation, while we were only able to partial dephosphorylate ATF1 in tissue lysates.
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All lanes : Anti-ATF1 antibody [EPR17028] (ab181569) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate treated with alkaline phosphatase for 1 hour at 37°C
Lysates/proteins at 25 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Developed using the ECL technique.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
ATF1 has been shown to be phosphorylated in cultured cells and hyperphosphorylation in tissues (PMID:20730097). Treatment of cell lysates with alkaline phosphatase lead to complete dephosphorylation, while we were only able to partial dephosphorylate ATF1 in tissue lysates.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling ATF1 with ab181569 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab181569 has been referenced in 2 publications.
- Zhao X et al. LINC00665/miR-9-5p/ATF1 is a novel axis involved in the progression of colorectal cancer. Hum Cell 33:1142-1154 (2020). PubMed: 32776307
- Tian J et al. Systematic Functional Interrogation of Genes in GWAS Loci Identified ATF1 as a Key Driver in Colorectal Cancer Modulated by a Promoter-Enhancer Interaction. Am J Hum Genet 105:29-47 (2019). PubMed: 31204011