RabMAb

Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292)

Overview

  • Product nameAnti-ATM (phospho S1981) antibody [EP1890Y]See all ATM primary antibodies ...
  • Description
    Rabbit monoclonal [EP1890Y] to ATM (phospho S1981)
  • Tested applicationsWB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, Rat
  • Immunogen

    A synthetic peptide corresponding to residues surrounding serine 1981 of human ATM was used as an immunogen.

  • Positive control
    • HEK293 cell lysate and Human gastric carcinoma.
  • General notes

    Produced under U.S. Patent No. 5,675,063. A 40 μl trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab81292 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 1/5000 - 1/10000. Detects a band of approximately 370 kDa (predicted molecular weight: 351 kDa).
IHC-P 1/100 - 1/250.
ICC/IF 1/100 - 1/250.
IP 1/40.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionSerine/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 specificityFound in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.
    • Involvement in diseaseDefects 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 similaritiesBelongs to the PI3/PI4-kinase family. ATM subfamily.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 1 PI3K/PI4K domain.
    • DomainThe 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 localizationNucleus. 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 complementation group A antibody
      • AT complementation group C antibody
      • AT complementation group D antibody
      • AT complementation group E antibody
      • AT mutated antibody
      • AT protein antibody
      • AT1 antibody
      • ATA antibody
      • Ataxia telangiectasia gene mutated in human beings 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_HUMAN antibody
      • DKFZp781A0353 antibody
      • Human phosphatidylinositol 3 kinase homolog antibody
      • MGC74674 antibody
      • OTTHUMP00000232981 antibody
      • Serine protein kinase ATM antibody
      • Serine-protein kinase ATM antibody
      • Serine/threonine-protein kinase ATM antibody
      • T cell prolymphocytic leukemia antibody
      • Tefu antibody
      • TEL1 antibody
      • TEL1, telomere maintenance 1, homolog antibody
      • TELO1 antibody
      • Telomere fusion protein antibody
      • TPLL antibody
      see all

    Anti-ATM (phospho S1981) antibody [EP1890Y] images

    • ab81292 staining ATM (phospho S1981) in Human primary fibroblasts treated with IR (5Gy) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 2% BSA for 2 hours at room temperature. Samples were incubated with primary antibody (1/500 in PBS + 2% BSA) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

      Left - ATM (phospho S1981), Right - DAPI + merge.

      See Abreview

    • ab81292 staining ATM in Human HeLa cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/250 in TBS-Tween20 + 5% BSA) for 1 hour at 22°C. A TRITC-conjugated Donkey anti-rabbit IgG polyclonal (1/150) was used as the secondary antibody. Cells treated with 10 Gy IR and allowed to recover for 30 minutes before fixed.

      See Abreview

    • ab81292, 1/100, staining Human gastric carcinoma by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    • All lanes : Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292) at 1/10000 dilution

      Lane 1 : HEK293 cell lysates untreated.
      Lane 2 : HEK293 cell lysates treated with doxorubicin.

      Lysates/proteins at 10 µg per lane.

      Secondary
      Lane 1 : HRP labelled goat anti-rabbit at 1/2000 dilution
      Lane 2 : HRP labelled goat anti-rabbit at 1/2000 dilution


      Predicted band size : 351 kDa
      Observed band size : 370 kDa (why is the actual band size different from the predicted?)
    • All lanes : Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292) at 1/1000 dilution

      Lane 1 : UV treated Human Lympocytes at 30 µg
      Lane 2 : UV treated Human Lympocytes at 20 µg
      Lane 3 : UV treated Human Lympocytes at 20 µg
      Lane 4 : UV treated Human Lympocytes at 20 µg

      Secondary
      Goat Polyclonal to Rabbit IgG at 1/2000 dilution
      developed using the ECL technique

      Performed under reducing conditions.

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


      Exposure time : 10 seconds

      Image courtesy Anonymous Abreview

      See Abreview

    • ICC/IF image of ab81292 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81292,1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • ab81292 showing positive staining in Breast carcinoma tissue.

    • ab81292 showing positive staining in Normal tonsil tissue.

    • ab81292 showing positive staining in Cervical carcinoma tissue.

    • ab81292 showing positive staining in Endometrial carcinoma tissue.

    References for Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292)

    This product has been referenced in:
    • Boichuk S  et al. Unbiased compound screening identifies unexpected drug sensitivities and novel treatment options for gastrointestinal stromal tumors. Cancer Res 74:1200-13 (2014). Read more (PubMed: 24385214) »
    • Chen W  et al. RIP1 maintains DNA integrity and cell proliferation by regulating PGC-1a-mediated mitochondrial oxidative phosphorylation and glycolysis. Cell Death Differ N/A:N/A (2014). Read more (PubMed: 24583643) »

    See all 33 Publications for this product

    Product Wall

    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa)
    Loading amount 30 µg
    Specification HeLa
    Treatment 10 Gy IR
    Gel Running Conditions Reduced Denaturing
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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    Submitted Apr 12 2013

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (4-12%)
    Sample Human Cell lysate - whole cell (lymphocytes)
    Specification lymphocytes
    Treatment UV treated
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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    Submitted Apr 07 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Sample Human Cell (Human primary fibroblasts treated with IR (5Gy))
    Specification Human primary fibroblasts treated with IR (5Gy)
    Permeabilization Yes - Yes - 0.2% Triton-X100
    Fixative Paraformaldehyde
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    Submitted Oct 11 2013

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Sample Human Cell (HeLa)
    Specification HeLa
    Permeabilization Yes - 0.1% Triton X-100
    Fixative Paraformaldehyde
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    Submitted Jul 04 2013

    Application Western blot
    Sample Human Cell lysate - whole cell (HCT116 colon cancer cell line)
    Loading amount 60 µg
    Specification HCT116 colon cancer cell line
    Treatment DMSO control and 10 µM etoposide for 24 hrs
    Gel Running Conditions Reduced Denaturing (7%)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Mr. Christian Marx

    Verified customer

    Submitted Oct 29 2012

    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa cells)
    Loading amount 25 µg
    Specification HeLa cells
    Treatment 10 Gy IR, 30 min recovery
    Gel Running Conditions Reduced Denaturing (4-16% Tris-Gly gel)
    Blocking step LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
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    Submitted Jan 10 2012

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (Blood lymphocytes)
    Specification Blood lymphocytes
    Fixative Paraformaldehyde
    Permeabilization Yes - 1% Triton-X
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
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    Submitted Sep 07 2011

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (lymphocytes)
    Specification lymphocytes
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.15% Triton X-100 for 10min
    Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 20°C
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    Submitted Feb 28 2011

    Application Western blot
    Sample Human Purified protein (Immunoprecipitated endogenous protein)
    Loading amount 0.1 µg
    Specification Immunoprecipitated endogenous protein
    Treatment Irradiated with 5Gy
    Gel Running Conditions Reduced Denaturing (4-12% Gradient Gel)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Oct 12 2010

    Application Western blot
    Sample Human Cell lysate - whole cell (A549 tumour cell line)
    Loading amount 50 µg
    Specification A549 tumour cell line
    Treatment 100IU Bleomycin 4h
    Gel Running Conditions Reduced Denaturing (3-8% tris acetate gel)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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    Submitted Jan 08 2010

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