Recombinant Anti-ATF2 antibody [E243] - ChIP Grade (ab32160)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E243] to ATF2 - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, IP, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ATF2 antibody [E243] - ChIP Grade
See all ATF2 primary antibodies -
Description
Rabbit monoclonal [E243] to ATF2 - ChIP Grade -
Host species
Rabbit -
Specificity
This antibody recognises ATF2, but does not cross react with other ATF family members. -
Tested applications
Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human ATF2 aa 450-550 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa cell lysate. IHC-P: Breast carcinoma tissue. Flow Cyt (intra): HeLa cells. IP: HeLa cell lysate. ChIP: Jurkat cells. ICC/IF: A549 cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Clonality
Monoclonal -
Clone number
E243 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32160 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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Flow Cyt (Intra) |
1/2000.
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ChIP |
Use 5 µg for 25 µg of chromatin.
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WB | (1) |
1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 54 kDa).
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IHC-P | (1) |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
1/60.
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ICC/IF |
1/250 - 1/500.
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Notes |
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Flow Cyt (Intra)
1/2000. |
ChIP
Use 5 µg for 25 µg of chromatin. |
WB
1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 54 kDa). |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
1/60. |
ICC/IF
1/250 - 1/500. |
Target
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Function
Transcriptional activator, probably constitutive, which binds to the cAMP-responsive element (CRE) (consensus: 5'-GTGACGT[AC][AG]-3'), a sequence present in many viral and cellular promoters. Interaction with JUN redirects JUN to bind to CRES preferentially over the 12-O-tetradecanoylphorbol-13-acetate response elements (TRES) as part of an ATF2/JUN complex. -
Tissue specificity
Abundant expression seen in the brain. -
Sequence similarities
Belongs to the bZIP family. ATF subfamily.
Contains 1 bZIP domain.
Contains 1 C2H2-type zinc finger. -
Post-translational
modificationsPhosphorylation of Thr-69 and Thr-71 by MAPK14 causes increased transcriptional activity. Also phosphorylated and activated by JNK. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 1386 Human
- Omim: 123811 Human
- SwissProt: P15336 Human
- Unigene: 592510 Human
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Alternative names
- Activating transcription factor 2 antibody
- Activating transcription factor 2 splice variant ATF2 var2 antibody
- ATF 2 antibody
see all
Images
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Anti-ATF2 antibody [E243] - ChIP Grade (ab32160) at 1/10000 dilution + HeLa cell lysate
Predicted band size: 54 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted? -
Ab32160, at a dilution of 1/250, staining ATF2 in paraffin embedded breast carcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunocytochemistry analysis of A549 (human lung carcinoma epithelial cell) labeling ATF2 with purified ab32160 at 1/100 dilution (10 µg/ml). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.10 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody. -
Chromatin was prepared from Jurkat (TPA and Ionomycin treated or not) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32160 (red), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
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ab32160 (purified) at 1/60 dilution (20 µg/mL) immunoprecipitating ATF2 in HeLa whole cell lysate.
Lane 3 (-): HeLa(Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µgab32160 & HeLa whole cell lysateRabbit monoclonal IgG (ab172730) instead of ab32160 in HeLa whole cell lysate
For western blotting, ab32160 at 1/1000 dilution (2.284 µg/mL) and
bbit TureBlot: Anti-Rabbit IgG HRP was used as the secondary antibody at 1/1500 dilution.
Blocking and diluting buffer: 5% NFDM /TBST . -
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATF2 (red) with ab32160 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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
References (8)
ab32160 has been referenced in 8 publications.
- Xu X et al. Increased ATF2 expression predicts poor prognosis and inhibits sorafenib-induced ferroptosis in gastric cancer. Redox Biol 59:102564 (2023). PubMed: 36473315
- Li ZJ et al. ATF2 accelerates the invasion and metastasis of hepatocellular carcinoma through targeting the miR-548p/TUFT1 axis. Hepatol Res 52:281-297 (2022). PubMed: 34904343
- Zu Y et al. The roles of CPSF6 in proliferation, apoptosis and tumorigenicity of lung adenocarcinoma. Aging (Albany NY) 14:9300-9316 (2022). PubMed: 36446361
- Huang C et al. Phosphoproteomic characterization of the signaling network resulting from activation of the chemokine receptor CCR2. J Biol Chem 295:6518-6531 (2020). PubMed: 32241914
- Liu J et al. The interplay between ATF2 and NEAT1 contributes to lung adenocarcinoma progression. Cancer Cell Int 20:594 (2020). PubMed: 33298086
- Hu Z et al. Transcriptional activation of miR-320a by ATF2, ELK1 and YY1 induces cancer cell apoptosis under ionizing radiation conditions. Int J Oncol 53:1691-1702 (2018). PubMed: 30066913
- Ma J et al. SPOP promotes ATF2 ubiquitination and degradation to suppress prostate cancer progression. J Exp Clin Cancer Res 37:145 (2018). PubMed: 29996942
- Wu X et al. CUG-binding protein 1 regulates HSC activation and liver fibrogenesis. Nat Commun 7:13498 (2016). WB ; Human . PubMed: 27853137