Overview

  • Product name

    Anti-ATM (phospho S1981) antibody [EP1890Y] - BSA and Azide free
    See all ATM primary antibodies
  • Description

    Rabbit monoclonal [EP1890Y] to ATM (phospho S1981) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, Dot blot, IHC-P, IPmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human ATM (phospho S1981).
    Database link: Q13315

  • Positive control

    • WB: HEK293 cell lysate treated with doxorubicin. IHC-P: Human gastric carcinoma, breast carcinoma, tonsil, cervical carcinoma, hepatocellular carcinoma and endometrial carcinoma tissues and mouse endometrium tissue. ICC/IF: HepG2 and HEK293 cells. IP: HEK293 cell lysate treated with doxorubicin.
  • General notes

    ab208775 is the carrier-free version of ab81292 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab208775 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208775 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
WB Use at an assay dependent concentration. Detects a band of approximately 371 kDa (predicted molecular weight: 351 kDa).

Please check the parent abID, ab81292, for more information on dilutions.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Dot blot Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      Serine/threonine protein kinase which activates checkpoint signaling upon double strand breaks (DSBs), apoptosis and genotoxic stresses such as ionizing ultraviolet A light (UVA), thereby acting as a DNA damage sensor. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX at double strand breaks (DSBs), thereby regulating DNA damage response mechanism. Also plays a role in pre-B cell allelic exclusion, a process leading to expression of a single immunoglobulin heavy chain allele to enforce clonality and monospecific recognition by the B-cell antigen receptor (BCR) expressed on individual B lymphocytes. After the introduction of DNA breaks by the RAG complex on one immunoglobulin allele, acts by mediating a repositioning of the second allele to pericentromeric heterochromatin, preventing accessibility to the RAG complex and recombination of the second allele. Also involved in signal transduction and cell cycle control. May function as a tumor suppressor. Necessary for activation of ABL1 and SAPK. Phosphorylates p53/TP53, FANCD2, NFKBIA, BRCA1, CTIP, nibrin (NBN), TERF1, RAD9 and DCLRE1C. May play a role in vesicle and/or protein transport. Could play a role in T-cell development, gonad and neurological function. Plays a role in replication-dependent histone mRNA degradation. Binds DNA ends.
    • Tissue specificity

      Found in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.
    • Involvement in disease

      Defects in ATM are the cause of ataxia telangiectasia (AT) [MIM:208900]; also known as Louis-Bar syndrome, which includes four complementation groups: A, C, D and E. This rare recessive disorder is characterized by progressive cerebellar ataxia, dilation of the blood vessels in the conjunctiva and eyeballs, immunodeficiency, growth retardation and sexual immaturity. AT patients have a strong predisposition to cancer; about 30% of patients develop tumors, particularly lymphomas and leukemias. Cells from affected individuals are highly sensitive to damage by ionizing radiation and resistant to inhibition of DNA synthesis following irradiation.
      Note=Defects in ATM contribute to T-cell acute lymphoblastic leukemia (TALL) and T-prolymphocytic leukemia (TPLL). TPLL is characterized by a high white blood cell count, with a predominance of prolymphocytes, marked splenomegaly, lymphadenopathy, skin lesions and serous effusion. The clinical course is highly aggressive, with poor response to chemotherapy and short survival time. TPLL occurs both in adults as a sporadic disease and in younger AT patients.
      Note=Defects in ATM contribute to B-cell non-Hodgkin lymphomas (BNHL), including mantle cell lymphoma (MCL).
      Note=Defects in ATM contribute to B-cell chronic lymphocytic leukemia (BCLL). BCLL is the commonest form of leukemia in the elderly. It is characterized by the accumulation of mature CD5+ B lymphocytes, lymphadenopathy, immunodeficiency and bone marrow failure.
    • Sequence similarities

      Belongs to the PI3/PI4-kinase family. ATM subfamily.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 1 PI3K/PI4K domain.
    • Domain

      The FATC domain is required for interaction with KAT5.
    • Post-translational
      modifications

      Phosphorylated by NUAK1/ARK5. Autophosphorylation on Ser-367, Ser-1893, Ser-1981 correlates with DNA damage-mediated activation of the kinase.
      Acetylation, on DNA damage, is required for activation of the kinase activity, dimer-monomer transition, and subsequent autophosphorylation on Ser-1981. Acetylated in vitro by KAT5/TIP60.
    • Cellular localization

      Nucleus. Cytoplasmic vesicle. Primarily nuclear. Found also in endocytic vesicles in association with beta-adaptin.
    • Information by UniProt
    • Database links

    • Alternative names

      • A-T mutated antibody
      • A-T mutated homolog antibody
      • AT mutated antibody
      • AT1 antibody
      • ATA antibody
      • Ataxia telangiectasia mutated antibody
      • Ataxia telangiectasia mutated gene antibody
      • Ataxia telangiectasia mutated homolog (human) antibody
      • Ataxia telangiectasia mutated homolog antibody
      • ATC antibody
      • ATD antibody
      • ATDC antibody
      • ATE antibody
      • ATM antibody
      • ATM serine/threonine kinase antibody
      • ATM_HUMAN antibody
      • DKFZp781A0353 antibody
      • MGC74674 antibody
      • OTTHUMP00000232981 antibody
      • Serine protein kinase ATM antibody
      • Serine-protein kinase ATM antibody
      • Serine/threonine-protein kinase ATM antibody
      • Tefu antibody
      • TEL1 antibody
      • TEL1, telomere maintenance 1, homolog antibody
      • TELO1 antibody
      • Telomere fusion protein antibody
      see all

    Images

    • Anti-ATM (phospho S1981) antibody [EP1890Y] - BSA and Azide free (ab208775) + HEK293 (human embryonic kidney) treated with Doxorubicin whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

      Predicted band size: 351 kDa
      Observed band size: 370 kDa
      why is the actual band size different from the predicted?


      Exposure time: 1 minute


      Blocking buffer and concentration: 5% NFDM/TBST

      Diluting buffer and concentration: 5% NFDM/TBST

    • ATM was immunoprecipitated from HEK-293 (Human embryonic kidney epithelial cell) treated with Doxorubicin whole cell lysate with ab81292 at 1/30 dilution (5 µg in 1 mg lysates). Western blot was performed from the immunoprecipitate using ab81292 at 1/2000 dilution. An anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
      Lane 1: HEK-293 treated with Doxorubicin whole cell lysate 10 µg (Input).
      Lane 2: ab81292 IP in HEK-293 treated with Doxorubicin whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab81292 in HEK-293 treated with Doxorubicin whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 minutes.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

    • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma) cells labeling ATM (phospho S1981) with purified ab81292 at 1/60 dilution (10 µg/mL) (red).

      Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

    • Dot blot analysis of ATM peptides using ab81292 at 1/000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody (ab97051) at 1/100,000 dilution.

      Blocking and diluting buffer was 5% NFDM/TBST, exposure time 3 minutes.

      Lane 1: ATM (pS1981) phospho peptide

      Lane 2: ATM non-phospho peptide

      Lane 3: ATM (pS428) phospho peptide

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labelling ATM (phospho S1981) with purified ab81292 at 1/70. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

      Negative control using PBS instead of primary antibody.

      Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue labeling ATM with unpurified ab81292 at a 1/100 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling ATM with unpurified ab81292.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil tissue labeling ATM with unpurified ab81292.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling ATM with unpurified ab81292.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrial carcinoma tissue labeling ATM with unpurified ab81292.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab81292).

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Sone K  et al. Critical role of lysine 134 methylation on histone H2AX for ?-H2AX production and DNA repair. Nat Commun 5:5691 (2014). Read more (PubMed: 25487737) »
    • Huang CY  et al. Distinct epidermal keratinocytes respond to extremely low-frequency electromagnetic fields differently. PLoS One 9:e113424 (2014). WB ; Human . Read more (PubMed: 25409520) »
    See all 2 Publications for this product

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