Overview

  • Product name
    Anti-ATR antibody - ChIP Grade
    See all ATR primary antibodies
  • Description
    Rabbit polyclonal to ATR - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein, corresponding to amino acids 400-460 of Human ATR.

  • Positive control
    • UV irradiated K562 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab2905 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
ICC/IF Use at an assay dependent dilution.
IP Use a concentration of 2 mg/ml.
WB 1/1000. Detects a band of approximately 305 kDa (predicted molecular weight: 309 kDa). (in lysate from UV irradiated K562 cells).
ChIP Use at an assay dependent dilution. PubMed: 20647539

Target

  • 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
    modifications
    Phosphorylated; 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
  • Database links
  • Alternative names
    • Ataxia telangiectasia and Rad3 related antibody
    • Ataxia telangiectasia and Rad3-related protein antibody
    • ATR antibody
    • ATR_HUMAN antibody
    • FCTCS antibody
    • FRAP Related Protein 1 antibody
    • FRAP-related protein 1 antibody
    • FRP1 antibody
    • MEC1 antibody
    • MEC1 mitosis entry checkpoint 1 homolog antibody
    • Protein kinase ATR antibody
    • RAC3 antibody
    • Rad3 related protein antibody
    • SCKL antibody
    • SCKL1 antibody
    • Serine/threonine protein kinase ATR antibody
    • Serine/threonine-protein kinase ATR antibody
    see all

Images

  • Western blot on ATR in K562 whole cell lysate using ab2905.
  • ab2905 staining ATR in the U2OS cell line from Human Osteosarcoma by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with Formaldehyde, permeabilized with NP-40 and blocked with 3% BSA for 60 minutes at 21°C. Samples were incubated with primary antibody (1/500 in PBS + 3% BSA) for 12 hour at 4°C. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal(1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-ATR antibody - ChIP Grade (ab2905) at 1/1000 dilution

    Lane 1 : MCF10A breast epithelial cells, grown in normoxic conditions (20% O2). Cells were lysed with NP40.
    Lane 2 : MCF10A Breast Epithelial cells grown in hypoxic. conditions (5% O2). Cells were lysed with NP40.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti Rabbit (HRP) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 309 kDa
    Observed band size: 300 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds



    Blocking: Carried out using non fat dry milk for 1 hr at room temperature.

    This image is an edited version of an image submitted courtesy of an Abreview by Francisco Ramirez-Valle submitted on 15th November 2005. We do not have any further information relating to this image.

    See Abreview

References

This product has been referenced in:
  • Li J & Zheng J Theaflavins prevent cartilage degeneration via AKT/FOXO3 signaling in vitro. Mol Med Rep 19:821-830 (2019). Read more (PubMed: 30569095) »
  • Lee SA  et al. INO80 haploinsufficiency inhibits colon cancer tumorigenesis via replication stress-induced apoptosis. Oncotarget 8:115041-115053 (2017). Read more (PubMed: 29383140) »
See all 22 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

That is really up to you, and what you want to do with the membrane: PVDF membranes tend to show a higher background, however, PVDF tends to be slightly more robust during handling and is easier to strip, whereas NC binds the protein stronger, but is more fragile during handling and not stable against organic solvents (if in contact by chance), and you can easier blot through.


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Question
Answer

Thank you for your enquiry. This antibody is not phospho-specific, but since it a polyclonal raised to a fusion protein using 60 amino acids of ATR, I would expect the antibody to have multiple epitopes and detect total ATR, including phosphorylated protein. However, I don't see that this has been specifically tested. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

We have received the following information regarding ab2905: It is best to try a high amp for about an hour. We recommend trying at 1 amp for 1 hour using appropriate methods to keep the system cool. We used 3 ml of 5% nonfat dried milk in TBST. Primary Ab. dilution of 1:1000 for 1 hour at room temp. Washed 3X with TBST buffer for 8 min at room temp. Each gently agitating solution. Secondary Ab 1:15000 in blocking buffer for 1 hour at room temp. Wash 3X with TBST for 8 min with agitation. Exposed for 15 seconds by ECL detection. We unfortunately are still waiting to hear regarding ab4471, my apologies for the delay,

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Answer

It has not been tested for its reactivity with ATM nor has the antibody been used in kinase assays, therefore we have no further information on these.

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