Anti-ATM antibody (ab82512)

Overview

• Product nameAnti-ATM antibodySee all ATM primary antibodies ...
• Description
  Rabbit polyclonal to ATM
• Tested applicationsWB, IP, ICC/IF more 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.

  Read Abcam's proprietary immunogen policy

  (Peptide available as ab90257.)

• Positive controlThis 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

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: 1% BSA, PBS, pH 7.4
• Concentration information loading...
• PurityImmunogen affinity purified
• Clonality Polyclonal
• IsotypeIgG
• Research Areas
  • Cancer
  • Oncoproteins/suppressors
  • Tumor suppressors
  • Other

  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • DNA Damage & Repair
  • DNA Damage Response
  • ATM / ATR

Applications

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
  modificationsPhosphorylated 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.

Target information above from: UniProt accession Q13315 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 395401 Chicken
  • Entrez Gene: 472 Human
  • Entrez Gene: 472 Human
  • Entrez Gene: 11920 Mouse
  • Omim: 607585 Human
  • SwissProt: Q13315 Human
  • SwissProt: Q13315 Human
  • SwissProt: Q62388 Mouse

  • Unigene: 367437 Human
  • Unigene: 5088 Mouse

  see all

• Alternative names
  A-T mutated antibodyA-T mutated homolog antibodyAT complementation group A antibody

  AT complementation group C antibodyAT complementation group D antibodyAT complementation group E antibodyAT mutated antibodyAT protein antibodyAT1 antibodyATA antibodyAtaxia telangiectasia gene mutated in human beings antibodyAtaxia telangiectasia mutated antibodyAtaxia telangiectasia mutated gene antibodyAtaxia telangiectasia mutated homolog (human) antibodyAtaxia telangiectasia mutated homolog antibodyAtaxia telangiectasia mutated protein antibodyATC antibodyATD antibodyATDC antibodyATE antibodyATM antibodyATM_HUMAN antibodyDKFZp781A0353 antibodyHuman phosphatidylinositol 3 kinase homolog antibodyMGC74674 antibodyOTTHUMP00000232981 antibodySerine protein kinase ATM antibodySerine-protein kinase ATM antibodySerine/threonine-protein kinase ATM antibodyT cell prolymphocytic leukemia antibodyTefu antibodyTEL1 antibodyTEL1 telomere maintenance 1 homolog antibodyTEL1, telomere maintenance 1, homolog antibodyTELO1 antibodyTelomere fusion protein antibodyTPLL antibody

  see all

Anti-ATM antibody images

• 

  Western blot - ATM antibody (ab82512)
  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
• 

  Western blot - ATM antibody (ab82512)
  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
• 

  Immunoprecipitation - Anti-ATM antibody (ab82512)
  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 70^oC; 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.
• 

  Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody (ab82512)
  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.