Recombinant
RabMAb

Anti-ATM antibody [Y170] (ab32420)

Overview

  • Product name
    Anti-ATM antibody [Y170]
    See all ATM primary antibodies
  • Description
    Rabbit monoclonal [Y170] to ATM
  • Specificity
    This antibody is specific for ATM.
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human ATM.
    (Peptide available as ab170988)

  • Positive control
    • WB: 293 cell lysate. FC: HeLa cells
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32420 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 1/1000 - 1/10000. Predicted molecular weight: 350 kDa.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.

 

Flow Cyt 1/20.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Serine/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 specificity
      Found in pancreas, kidney, skeletal muscle, liver, lung, placenta, brain, heart, spleen, thymus, testis, ovary, small intestine, colon and leukocytes.
    • Involvement in disease
      Defects 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 similarities
      Belongs to the PI3/PI4-kinase family. ATM subfamily.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 1 PI3K/PI4K domain.
    • Domain
      The 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 localization
      Nucleus. 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 mutated antibody
      • AT1 antibody
      • ATA 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
      • MGC74674 antibody
      • OTTHUMP00000232981 antibody
      • Serine protein kinase ATM antibody
      • Serine-protein kinase ATM antibody
      • Serine/threonine-protein kinase ATM antibody
      • Tefu antibody
      • TEL1 antibody
      • TEL1, telomere maintenance 1, homolog antibody
      • TELO1 antibody
      • Telomere fusion protein antibody
      see all

    Images

    • All lanes : Anti-ATM antibody [Y170] (ab32420) at 1/3000 dilution (Purified)

      Lane 1 : HeLa cell lysate
      Lane 2 : HEK293 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 350 kDa
      Observed band size : 370 kDa (why is the actual band size different from the predicted?)
    • Formaldehyde-fixed human serous ovarian tumor tissue stained for ATM using ab32420 at 1/50 dilution in immunohistochemical analysis.

      See Abreview

    • Formaldehyde-fixed human colon tissue stained for ATM using ab32420 at 1/100 dilution in immunohistochemical analysis.

      See Abreview

    • Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ATM with ab32420 at 1:100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.

    • Immunofluorescent staining of HeLa cells (fixed with 4% PFA) with purified ab32420 at a dilution of 1/250. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.

    • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling ATM with purified ab32420 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Immunohistochemical analysis of paraffin-embedded Human normal breast tissue labeling ATM with ab32420 at 1:100 dilution. Tissue underwent antigen retrieval using Tris/EDTA Buffer (pH9.0). The section was counterstained with haematoxylin.

    References

    This product has been referenced in:
    • Golia B  et al. ATM induces MacroD2 nuclear export upon DNA damage. Nucleic Acids Res 45:244-254 (2017). Read more (PubMed: 28069995) »
    • Royba E  et al. Evaluation of ATM heterozygous mutations underlying individual differences in radiosensitivity using genome editing in human cultured cells. Sci Rep 7:5996 (2017). Read more (PubMed: 28729543) »

    See all 44 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 20% · Temperature: RT°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA Buffer
    Sample
    Human Tissue sections (Serous Ovarian Tumour)
    Specification
    Serous Ovarian Tumour
    Permeabilization
    No
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jul 15 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
    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

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Colon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization
    No
    Specification
    Colon
    Blocking step
    hydrogen peroxide as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    Formaldehyde
    Username

    Vinicius Kannen Cardoso

    Verified customer

    Submitted Aug 10 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Colon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH9 20 mins Leica EDTA
    Permeabilization
    No
    Specification
    Colon
    Blocking step
    (agent) for 5 minute(s) · Concentration: 10%
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Feb 29 2016

    The blocking peptide for the Anti-ATM antibody [Y170] (ab32420) has now been added to the catalogue following your enquiry. This can be accessed under the catalogue number ab170988


    For producing the IHC-P images, including the glioma tissue, on the datasheet for ab32420, we used the following protocol with a sodium citrate heat mediated antigen retrieval:
    http://www.epitomics.com/products/support/ihc
    We had success w...

    Read More

    Gracias por tu respuesta.
    Efectivamente, si haces un pedido de ab26, el ab32420 os saldría gratuito, al entrar en la promoción de RabMabs.
    Simplemente indica el código promocional RABMAB-XBSMG en la casilla destinada para ello cuando realizas...

    Read More

    Thank you for contacting us.
    Unfortunately I cannot find this particular lot number for this product. I have checked the orders from your Organisation for this antibody and found an order from December 2011. The lot number for the vial sent is XXX...

    Read More

    Thank you for contacting us and your interest in our products.

    Epitomics did offer isotype controls specific for different lots of antibody purchased. This was providing the antibody at the same concentration as the antibody to be tested. We d...

    Read More

    Thank you for contacting Abcam.

    The peptides for both ab32420 and ab81292 maps between amino acids ***********of human ATM (UniProt: Q13315).

    Please let me know if there is anything else I can help you with.

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

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