Overview

  • Product nameAnti-ATM antibody [2C1 (1A1)]
    See all ATM primary antibodies
  • Description
    Mouse monoclonal [2C1 (1A1)] to ATM
  • Conjugation notes-
  • SpecificityThe ATM antibody, clone 2C1, recognizes full-length ATM.
  • Tested applicationsFlow Cyt, ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey
  • Immunogen

    Fusion protein expressed in E. coli corresponding to amino acids 2577-3056.

  • Positive control
    • lymphoblastoid nuclear lysate, human Raji, U87-MG , SK-N-SH (human neuroblastoma), IMR32 , SK-N-AS.

Properties

Applications

Our Abpromise guarantee covers the use of ab78 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 1-2µg for 106 cells.
ICC/IF Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml.
WB 1/2000. Detects a band of approximately 350 kDa (predicted molecular weight: 350 kDa).
IP Use a concentration of 1 - 10 µg/ml.

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
    • 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 antibody [2C1 (1A1)] images

  • ab78 staining ATM in Human U2OS cells by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 0.5% NP40/PBS and blocked with 3% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/100 in 3% BSA/PBS) for 12 hours at 4°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/200) (ab150077) was used as the secondary antibody.

  • All lanes : Anti-ATM antibody [2C1 (1A1)] (ab78) at 1/2000 dilution

    Lane 1 : 50ug lymphoblastoid nuclear lysate, anti-beta actin antibody.
    Lane 2 : 25ug lymphoblastoid nuclear lysate, anti-beta actin antibody.
    Lane 3 : No ATM negative control, anti-beta actin antibody.

    Secondary
    Anti-mouse secondary antibody at 1/2000 dilution

    Performed under reducing conditions.

    Predicted band size : 350 kDa

    Simona Cavalieri, University of Turin, Italy

    Arrows denote the location of the 45kDa beta actin protein, and the 350kDa ATM protein.


  • Predicted band size : 350 kDa

    Detection of human ATM protein using anti-ATM 2C1 monoclonal antibody (ab78) by western blot.

  • Detection of human ATM protein using anti-ATM 2C1 monoclonal antibody (ab78) by immunoprecipitation.



  • Predicted band size : 350 kDa

    Detection of ATM (Anti-ATM 2C1, ab78) or ATR (Anti-ATR 2B5, ab4471) by western blot in human Raji whole cell extract.

  • ab78 (2µg/ml) staining ATM in human colonic mucosa, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining of mucosal epithelium and lymphocytes.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

  • Overlay histogram showing HeLa cells stained with ab78 (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 (ab78, 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 antibody [2C1 (1A1)] (ab78)

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) »
  • Oleson BJ  et al. Nitric oxide induces ataxia telangiectasia mutated (ATM) protein-dependent ?H2AX protein formation in pancreatic ß cells. J Biol Chem 289:11454-64 (2014). WB ; Rat . Read more (PubMed: 24610783) »

See all 19 Publications for this product

Product Wall

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Sample Human Cell lysate - nuclear (HeLa nuclear cell lysate)
Specification HeLa nuclear cell lysate
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 Nov 20 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Treatment Doxorubicin for 3 hours
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Andrew Cobb

Verified customer

Submitted Jul 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2OS)
Specification U2OS
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% NP40/PBS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 27 2012

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab78with the order number 1009642. To check the status of the order p...

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Thank you for your enquiry. The Abcam lab runs Invitrogen pre-cast SDS-PAGE gels at 200 volts for 35mins, and runs the transfer for 60 mins at 30 volts. I hope this information helps, please do not hesitate to contact us if you need any more advi...

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Thank you for getting back to me. I am sorry for my lack of clarity. I would like to recommend you perform a blocking step using 3% (or 5%) BSA in TBST for 1 hour and perform antibody incubations with the antibody diluted in TBST ovenight at 4oC. We...

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Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. ATM antibody [2C1 (1A1)] (ab78) is a relatively good selling antibody that we have received some recent positive feedback for. I have read you...

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