Recombinant Anti-ATR (phospho S428) antibody [EPR2184] (ab178407)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2184] to ATR (phospho S428)
- Suitable for: Dot blot, IHC-P, WB
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ATR (phospho S428) antibody [EPR2184]
See all ATR primary antibodies -
Description
Rabbit monoclonal [EPR2184] to ATR (phospho S428) -
Host species
Rabbit -
Specificity
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
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Tested applications
Suitable for: Dot blot, IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC: Human breast tissue, human thyroid tissue; WB: Untreated HeLa, HeLa treated with 4mM hydroxyurea for 20 hours whole cell 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
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2184 -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab178407 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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Dot blot |
1/1000.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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WB |
1/1000 - 1/10000. Predicted molecular weight: 300 kDa.
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Notes |
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Dot blot
1/1000. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
WB
1/1000 - 1/10000. Predicted molecular weight: 300 kDa. |
Target
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Function
Serine/threonine protein kinase which activates checkpoint signaling upon genotoxic stresses such as ionizing radiation (IR), ultraviolet light (UV), or DNA replication stalling, thereby acting as a DNA damage sensor. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates BRCA1, CHEK1, MCM2, RAD17, RPA2, SMC1 and p53/TP53, which collectively inhibit DNA replication and mitosis and promote DNA repair, recombination and apoptosis. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX at sites of DNA damage, thereby regulating DNA damage response mechanism. Required for FANCD2 ubiquitination. Critical for maintenance of fragile site stability and efficient regulation of centrosome duplication. -
Tissue specificity
Ubiquitous, with highest expression in testis. Isoform 2 is found in pancreas, placenta and liver but not in heart, testis and ovary. -
Involvement in disease
Defects in ATR are a cause of Seckel syndrome type 1 (SCKL1) [MIM:210600]. SCKL1 is a rare autosomal recessive disorder characterized by growth retardation, microcephaly with mental retardation, and a characteristic 'bird-headed' facial appearance. -
Sequence similarities
Belongs to the PI3/PI4-kinase family. ATM subfamily.
Contains 1 FAT domain.
Contains 1 FATC domain.
Contains 2 HEAT repeats.
Contains 1 PI3K/PI4K domain. -
Post-translational
modificationsPhosphorylated; autophosphorylates in vitro. -
Cellular localization
Nucleus. Nucleus > PML body. Depending on the cell type, it can also be found in PML nuclear bodies. Recruited to chromatin during S-phase. Redistributes to discrete nuclear foci upon DNA damage, hypoxia or replication fork stalling. - Information by UniProt
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Database links
- Entrez Gene: 545 Human
- Omim: 601215 Human
- SwissProt: Q13535 Human
- Unigene: 271791 Human
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Alternative names
- Ataxia telangiectasia and Rad3 related antibody
- Ataxia telangiectasia and Rad3-related protein antibody
- ATR antibody
see all
Images
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Immunohistochemical analysis of Paraffin-embedded human thyroid tissue sections labelling ATR with ab178407 at 1/5000 dilution followed by ready to use secondary ab209101 Rabbit specific IHC polymer detection kit HRP/DAB. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Nuclear staining on human thyroid without alkaline phosphatase (or Lambda Protein Phosphatase) treatment; No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab178407 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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Immunohistochemical analysis of Paraffin-embedded human breast tissue sections labelling ATR with ab178407 at 1/5000 dilution followed by ready to use secondary ab209101 Rabbit specific IHC polymer detection kit HRP/DAB. Counterstained with Hematoxylin. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
Nuclear staining on human breast without alkaline phosphatase (or Lambda Protein Phosphatase) treatment; No signal was detected when tissues were treated with alkaline phosphatase (image B). The section was incubated with ab178407 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument
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All lanes : Anti-ATR (phospho S428) antibody [EPR2184] (ab178407) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa treated with 4mM hydroxyurea for 20 hours whole cell lysate
Lane 3 : HeLa treated with 4mM hydroxyurea for 20 hours whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 300 kDa
Observed band size: 270 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST
Exposure time: 60 seconds
We are unsure of the nature of the 160kDa band.
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Dot blot analysis using 1/1000 dilution ab178407 and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary at 1/100000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST
Lane 1: ATR non-phospho peptide
Lane 2: ATR S428 phospho peptide
Lane 3: ATR S435 phospho peptide
Lane 4: ATR S428+S435 phospho peptide
Exposure time: 3 minutes
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (18)
ab178407 has been referenced in 18 publications.
- Pádua JDB et al. Prognostic Value of the Immunohistochemical Expression of RAD51 and BRCA2 in Gastric Adenocarcinoma. J Histochem Cytochem 70:199-210 (2022). PubMed: 34978208
- Hwang HJ et al. Ginsenoside Rh2 sensitizes the anti-cancer effects of sunitinib by inducing cell cycle arrest in renal cell carcinoma. Sci Rep 12:19752 (2022). PubMed: 36396667
- Zhou S et al. MET inhibition enhances PARP inhibitor efficacy in castration-resistant prostate cancer by suppressing the ATM/ATR and PI3K/AKT pathways. J Cell Mol Med 25:11157-11169 (2021). PubMed: 34761497
- Cai T et al. Notoginsenoside R1 induces DNA damage via PHF6 protein to inhibit cervical carcinoma cell proliferation. Mol Med Rep 23:N/A (2021). PubMed: 33537816
- Rudd SG et al. MTH1 Inhibitor TH588 Disturbs Mitotic Progression and Induces Mitosis-Dependent Accumulation of Genomic 8-oxodG. Cancer Res 80:3530-3541 (2020). PubMed: 32312836