Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)

Overview

  • Product nameAnti-ATM (phospho S1981) antibody [10H11.E12]
    See all ATM primary antibodies
  • Description
    Mouse monoclonal [10H11.E12] to ATM (phospho S1981)
  • Specificityab36810 is specific for the human ATM kinase.
  • Tested applicationsFlow Cyt, ICC/IF, WB, ICC, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (Human) with a phosphorylated serine (1981)

  • Positive control
    • Irradiated normal Human fibroblasts (no reactivity against non-irradiated cell extracts).

Properties

Applications

Our Abpromise guarantee covers the use of ab36810 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
Flow Cyt Use 1µg for 106 cells.
ICC/IF Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 370 kDa (predicted molecular weight: 370 kDa).

Abcam recommends using 3% Milk as the blocking agent.

ICC 1/1000.
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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 [10H11.E12] images

  • ab36810 staining ATM (phospho S1981) in T98G Human brain glioblastoma cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, pH 7.4 for 5 minutes at room temperature and blocked with 5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/250 in PBS) for 1 hour. A CF488A-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810) at 10 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Extract from Patient with Ataxia-Telangiectasia Whole Cell Lysate
    Lane 3 : Irradiated HeLa Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under non-reducing conditions.

    Predicted band size : 370 kDa
    Observed band size : 370 kDa
    Additional bands at : 100 kDa,110 kDa,145 kDa,200 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • All lanes : Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810) at 1/1000 dilution

    Lane 1 : Control lane
    Lane 2 : Irradiated Human fibroblasts (10 Gy gamma-irradiation)
    Lane 3 : Molecular weight marker
    Lane 4 : Peroxidated Human fibroblasts (300 µM hydrogen peroxide)
    Lane 5 : Peroxidated Human fibroblasts (1 mM hydrogen peroxide)
    Lane 6 : Peroxidated Human fibroblasts (10 mM hydrogen peroxide)

    developed using the ECL technique

    Predicted band size : 370 kDa
    Observed band size : 370 kDa
    Lysate loading concentration: 40µg
  • Ab36810staining Human colon. Staining is localised to the nucleus.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the DAKO 3-in-1 antigen retrieval buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • Overlay histogram showing HeLa cells stained with ab36810 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab36810, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)

This product has been referenced in:
  • Morgenroth A  et al. Breaking the invulnerability of cancer stem cells: two-step strategy to kill the stem-like cell subpopulation of multiple myeloma. Mol Cancer Ther 13:144-53 (2014). Read more (PubMed: 24174494) »
  • Schuler N  et al. DNA-damage foci to detect and characterize DNA repair alterations in children treated for pediatric malignancies. PLoS One 9:e91319 (2014). ICC ; Human . Read more (PubMed: 24637877) »

See all 11 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample Human Cell (T98G Human brain glioblastoma)
Specification T98G Human brain glioblastoma
Permeabilization Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative Paraformaldehyde
Username

Dr. Dimitra Kalamida

Verified customer

Submitted Dec 12 2013

Thank you for contacting Abcam.

I am sorry about the issues you have been having with ab36818. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples. Since the current lot is not...

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I have another lot available. Even though this is a monoclonal antibody, it may make a difference. I would be happy to send another vial to you. Please let me know if you are agreeable to this or if you would p...

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I am not sure why you are obtaining such a high degree of background with this antibody. Since your other blot is very clean, it is clearly not a technique issue such as insufficient washing. <...

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I am sorry that you have been experiencing problems with this antibody. I understand you are still troubleshooting based on the protocol advice I provided. Please do not hesit...

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I am sorry to hear that you are still experiencing difficulties with this antibody in WB. I have reviewed the protocol modifications you made and the resulting blots. One thing I am not clear about is the amoun...

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Thank you for contacting Abcam regarding ab36810.


I am sorry that you have been experiencing difficulties with this antibody and detecting a high level of background. I have reviewed the protocol and images you have provided and I would...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has...

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Thank you for this further information.

I am sorry that this product has not performed as stated on the datasheet. This product is guaranteed to work in WB on rat and is covered by our Abpromise for a period of six months. Since we have been ...

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