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 applicationsSuitable for: WB, IHC-P, IP, Flow Cytmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human ATM (phospho S1981).

  • 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. IP: HEK293 cell lysate treated with doxorubicin. FC: HepG2 cells
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

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

     

    Alternative versions available:

    Anti-ATM (phospho S1981) antibody (HRP) [EP1890Y] (ab193607)

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 1/50000. Detects a band of approximately 370 kDa (predicted molecular weight: 351 kDa).

For unpurified use at 1/5000 - 1/10000.

IHC-P 1/70 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

IP 1/30 - 1/40.

 

Flow Cyt 1/60.
  • Application notesIs unsuitable for ICC/IF.
  • 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 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

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

    • All lanes : Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292) at 1/50000 dilution (purified)

      Lane 1 : HEK293 cell lysate, untreated
      Lane 2 : HEK293 cell lysate, treated with Doxorubicin

      Lysates/proteins at 10 µg per lane.

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

      Blocking and dilution buffer: 5% NFDM/TBST.

    • 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, a 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.

    • All lanes : Anti-ATM (phospho S1981) antibody [EP1890Y] (ab81292) at 1/10000 dilution (unpurified)

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

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution

      Predicted band size : 351 kDa
      Observed band size : 370 kDa (why is the actual band size different from the predicted?)
    • Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling ATM (phospho S1981) with purified ab81292 at 1/60 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised 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.

    • 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 1/100000 dilution. Blocking and diluting buffer was 5% NFDM/TBST, exposure time 3 minuts.

       

      Lane 1: ATM (pS1981) phospho peptide

      Lane 2: ATM non-phospho peptide

      Lane 3: ATM (pS428) phospho peptide

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

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

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

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

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

    • ab81292 (purified) at 1/30 immunoprecipitating RAB27A in 293 + doxorubicin whole cell lysate, 10ug (Lane 1). Lane 2 - Rabbit monoclonal IgG (ab172730) instead of ab81292 in HeLa whole cell lysate. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

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

    This product has been referenced in:
    • Franz A  et al. Chromatin-associated degradation is defined by UBXN-3/FAF1 to safeguard DNA replication fork progression. Nat Commun 7:10612 (2016). Read more (PubMed: 26842564) »
    • García CP  et al. Human Pluripotent Stem Cells and Derived Neuroprogenitors Display Differential Degrees of Susceptibility to BH3 Mimetics ABT-263, WEHI-539 and ABT-199. PLoS One 11:e0152607 (2016). ICC/IF ; Human . Read more (PubMed: 27030982) »

    See all 59 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    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
    Username

    Abcam user community

    Verified customer

    Submitted Apr 12 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (Colon)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization Yes - tween 20
    Specification Colon
    Blocking step hydrogen peroxide as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative Formaldehyde
    Username

    Vinicius Kannen Cardoso

    Verified customer

    Submitted Aug 10 2016




    I can confirm that HEK293 cells were treated with 10uM doxorubicin for 24 hours. After the treatment, the cells were prepared into lysates for WB as normal.


    We offer Doxorubicin in two sizes, 10 and 50 mg:


    ab1...

    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (U2OS cells)
    Permeabilization Yes - 0.25% Triton
    Specification U2OS cells
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Fixative Paraformaldehyde
    Username

    Dr. Remi Buisson

    Verified customer

    Submitted Nov 21 2014

    Application Western blot
    Loading amount 10 µg
    Gel Running Conditions Reduced Denaturing (6)
    Sample Human Cell lysate - whole cell (Hela cells)
    Specification Hela cells
    Treatment Irradiation for 15 min
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 24 2014

    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
    Username

    Abcam user community

    Verified customer

    Submitted Apr 07 2014

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (6%)
    Sample Human Cell lysate - whole cell (HCT116 colon carcinoma)
    Specification HCT116 colon carcinoma
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 24 2014

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

    Abcam user community

    Verified customer

    Submitted Oct 11 2013

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

    Abcam user community

    Verified customer

    Submitted Jul 04 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    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
    Username

    Mr. Christian Marx

    Verified customer

    Submitted Oct 29 2012

    1-10 of 15 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"