Overview

  • Product nameAnti-ATM antibodySee all ATM primary antibodies ...
  • Description
    Rabbit polyclonal to ATM
  • Tested applicationsWB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chicken, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 3000 to the C-terminus of Human ATM.

    (Peptide available as ab90257.)

  • Positive control
    • This antibody gave a positive signal in the following cell lysates: HeLa whole cell; HeLa nuclear; HeLa whole cell - irradiated; HepG2, HEK293, NIH3T3.UV treated HeLa cell line (IF).

Properties

Applications

Our Abpromise guarantee covers the use of ab82512 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 Use a concentration of 1 µg/ml. Detects a band of approximately 320 kDa (predicted molecular weight: 350 kDa).Can be blocked with ATM peptide (ab90257).
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 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
    • 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
    • Ataxia telangiectasia mutated protein 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
    • TEL1, telomere maintenance 1, homolog antibody
    • TELO1 antibody
    • Telomere fusion protein antibody
    • TPLL antibody
    see all

Anti-ATM antibody images

  • All lanes : Anti-ATM antibody (ab82512) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 350 kDa
    Observed band size : 320 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 195 kDa (possible isoform).

    Exposure time : 15 minutes
  • All lanes : Anti-ATM antibody (ab82512) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : HeLa Whole Cell Lysate - Irradiated (5Gy)
    Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 350 kDa
    Observed band size : 350 kDa
    Additional bands at : 160 kDa,195 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • ATM was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to ATM and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab82512.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: Bands: 320kDa: ATM; 195kDa: We believe that the band observed at 195 kDa corresponds to isoform 2.
  • ICC/IF image of ab82512 stained HeLa cells. The cells were exposed to 2000µJ/cm2 of UV and incubated at 37°C for 24 hours and then were 4% formaldehyde 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 (ab82512, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) 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.

References for Anti-ATM antibody (ab82512)

ab82512 has not yet been referenced specifically in any publications.

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
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Dr. Dimitra Kalamida

Verified customer

Submitted Dec 12 2013

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