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
  • 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. ICC/IF: HepG2 and HEK293 cells. IP: HEK293 cell lysate treated with doxorubicin.
  • General notes

    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 40 µl 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. 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).

For unpurified use at 1/100 - 1/250.

ICC/IF 1/70.

For unpurified use at 1/100 - 1/250.

IP 1/30.

For unpurified use at 1/40.

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 serine/threonine kinase 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

  • 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.

  • 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?)
  • Unpurified 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. A goat anti rabbit Alexa Fluor® 488 (IgG polyclonal; ab150077 (1/1000) was used as the secondary antibody.

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

    See Abreview

  • Unpurified 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

  • ICC/IF image of unpurified 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 (unpurified ab81292,1µg/ml) overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (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.

  • 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse endometrium 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.

  • 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:
  • 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 41 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

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

Abcam user community

Verified customer

Submitted Oct 11 2013

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

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

Abcam user community

Verified customer

Submitted Jan 10 2012

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

Abcam user community

Verified customer

Submitted Sep 07 2011

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

Abcam user community

Verified customer

Submitted Feb 28 2011

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