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Anti-ATM antibody [Y170] ab32420 is a rabbit monoclonal antibody that is used in ATM western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone Y170 has been tried and trusted by researchers since 2006 and is cited in >170 publications
- Specificity confirmed with ATM knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation


Images

Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420), expandable thumbnail
  • Western blot - Anti-ATM antibody [Y170] (AB32420), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (AB32420), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (AB32420), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Not recommended
Tested
Tested
Tested
Tested

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

-

Tested
Tested

Species

Human

Dilution info

1/20

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

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Target data

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 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 DYRK2, CHEK2, p53/TP53, FANCD2, NFKBIA, BRCA1, CTIP, nibrin (NBN), TERF1, UFL1, RAD9, UBQLN4 and DCLRE1C (PubMed:9843217, PubMed:9733515, PubMed:10550055, PubMed:10766245, PubMed:10839545, PubMed:10910365, PubMed:10802669, PubMed:10973490, PubMed:11375976, PubMed:12086603, PubMed:15456891, PubMed:19965871, PubMed:30612738, PubMed:30886146). 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. Phosphorylation of DYRK2 in nucleus in response to genotoxic stress prevents its MDM2-mediated ubiquitination and subsequent proteasome degradation. Phosphorylates ATF2 which stimulates its function in DNA damage response. Phosphorylates ERCC6 which is essential for its chromatin remodeling activity at DNA double-strand breaks (PubMed:29203878).

Alternative names

Serine-protein kinase ATM, Ataxia telangiectasia mutated, A-T mutated, ATM

Recommended products

Anti-ATM antibody [Y170] ab32420 is a rabbit monoclonal antibody that is used in ATM western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone Y170 has been tried and trusted by researchers since 2006 and is cited in >170 publications
- Specificity confirmed with ATM knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

Y170

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

ATM also known as Ataxia Telangiectasia Mutated is a protein kinase with a molecular weight of approximately 370 kDa. ATM protein primarily resides in the cell nucleus and functions as a critical regulator of the cell cycle. It plays a significant role in the detection of DNA damage and initiation of repair processes. As part of its mechanical functions ATM phosphorylates serine and threonine residues on various substrates most notably in response to double-strand breaks in DNA. This activity is important for maintaining genomic stability.

Biological function summary

ATM acts as a coordinator in cellular response to DNA damage highly interacting with multiple components of the DNA repair machinery. It forms a complex with proteins like NBS1 and MRN complex facilitating repair by recruiting and activating other proteins involved in homologous recombination and non-homologous end joining pathways. ATM also modulates p53 activity a primary response factor in cellular stress management linking ATM to control of cell cycle arrest and apoptosis. This positions ATM as an integral part of maintaining cellular integrity in face of genomic insult.

Pathways

ATM integrates neatly within the DNA damage response and cell cycle control pathways. ATM's operative relationship with the MRN complex and its role in the PI3K-related protein kinase family helps initiate appropriate repair processes upon DNA damage detection. Additionally ATM regulates the activity of proteins such as Chk2 which further propagates signals to p53 influencing decisions between cell cycle arrest and apoptosis. These interactions link ATM closely to essential processes like DNA repair and cell survival highlighting its role in genomic maintenance.

Associated diseases and disorders

ATM mutations or dysregulation leads to Ataxia Telangiectasia an autosomal recessive disorder characterized by neurodegeneration immune deficiencies and cancer predisposition. ATM dysfunction also connects to cancer development particularly breast cancer where it transmits signals involving BRCA1 contributing to DNA repair through homologous recombination. Understanding ATM dynamics and related pathways has important implications for developing therapeutic strategies to manage or mitigate effects associated with its dysfunction.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.

  • Western blot - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Western blot - Anti-ATM antibody [Y170] (ab32420)

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

    Lane 1: HeLa cell lysate at 10 µg

    Lane 2: HEK293 cell lysate at 10 µg

    Secondary

    All lanes: HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 351 kDa

    Observed band size: 370 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420)

    Immunocytochemistry/ Immunofluorescence analysis of HepG2 cells labeling ATM with ab32420 at 1/500. Goat anti rabbit IgG(Alexa Fluor® 488), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/1000 was used as the secondary antibody. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Nulei were counterstained blue with DAPI.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail
    Image from Adeyemi R.O. et al PLoS Pathog. 2010 Oct 7;6(10):e1001141. doi: 10.1371/journal.ppat.1001141.

    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [Y170] (ab32420)

    DNA repair proteins accumulate at MVM APAR bodies

    Repair proteins accumulate at APAR bodies. NB324K cells were infected with MVMp (MOI of 10) for 16 hr before being fixed and processed for immunofluorescence. Cells were stained with the indicated antibodies to mark DDR repair proteins. APAR bodies were detected with antibodies to NS1. Nuclei were stained with DAPI. All images were captured using an objective of 63×.

    Cells were fixed with 4% paraformaldehyde for 15 minutes and permeabilized with 0.5% Triton X-100 in PBS for 15 minutes.

    (Image shows the right-hand panel of Figure 2A)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420)

    Formaldehyde-fixed human serous ovarian tumor tissue stained for ATM using ab32420 at 1/50 dilution in immunohistochemical analysis.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420)

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

  • Western blot - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Western blot - Anti-ATM antibody [Y170] (ab32420)

    False colour image of Western blot: Anti-ATM antibody [Y170] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32420 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type A549 cell lysates with no signal observed at this size in ATM knockout cell line Human ATM knockout A549 cell line ab276095 (knockout cell lysate ab283834). To generate this image, wild-type and ATM knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-ATM antibody [Y170] (ab32420) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: ATM knockout A549 cell lysate at 20 µg

    Lane 3: HEK-293 cell lysate at 20 µg

    Lane 4: U-2 OS cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 351 kDa

    Observed band size: 350 kDa

  • Flow Cytometry (Intracellular) - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ATM antibody [Y170] (ab32420)

    Intracellular 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 Fluorr®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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATM antibody [Y170] (ab32420)

    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.

  • Western blot - Anti-ATM antibody [Y170] (ab32420), expandable thumbnail

    Western blot - Anti-ATM antibody [Y170] (ab32420)

    Western blot: Anti-ATM antibody [Y170] (ab32420) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32420 was shown to bind specifically to ATM. A band was observed at 350 kDa in wild-type MCF7 cell lysates with a reduction in signal observed at this size in ATM heterozygous knockout cell line Human ATM heterozygous knockout MCF7 cell line ab282630. To generate this image, wild-type and ATM heterozygous knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-ATM antibody [Y170] (ab32420) at 1/1000 dilution

    Lane 1: Wild-type MCF7 cell lysate at 20 µg

    Lane 2: ATM knockout MCF7 cell lysate at 20 µg

    Lane 3: Wild-type A549 cell lysate at 20 µg

    Lane 4: ATM knockout A549 ab283811 cell lysate at 20 µg

    Observed band size: 350 kDa

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Product protocols

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