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Rabbit Recombinant Monoclonal ATM antibody. Carrier free. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

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Images

Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (AB223533), expandable thumbnail
  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (AB223533), expandable thumbnail
  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (AB223533), expandable thumbnail
  • ChIP - Anti-ATM antibody [EPR20100] - BSA and Azide free (AB223533), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (AB223533), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Rat
Expected
Expected
Tested
Expected
Expected

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

7 products for Alternative Product

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

Rabbit Recombinant Monoclonal ATM antibody. Carrier free. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Yes

Clone number

EPR20100

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab223533 is the carrier-free version of Anti-ATM antibody [EPR20100] - ChIP Grade ab201022.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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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9 product images

  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    False colour image of Western blot: Anti-ATM antibody [EPR20100] - ChIP Grade 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, Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 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 [EPR20100] - ChIP Grade (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022) 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

  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 3 minutes; Lane 3: 30 seconds; Lane 4: 5 seconds; Lane 5: 1 second

    All lanes: Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533) at 1/1000 dilution

    Lane 1: Human testis lysate at 10 µg

    Lane 2: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 3: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

    Lane 4: 293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

    Lane 5: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 351 kDa

    Observed band size: 351 kDa

  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533) at 1/5000 dilution

    All lanes: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 351 kDa

    Observed band size: 351 kDa

    Exposure time: 10s

  • ChIP - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    ChIP - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    Chromatin was prepared from HCT 116 (Human colorectal carcinoma cell line) cells treated with 1mM Hydroxyurea for 16h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow).  The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATM with Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

  • Immunoprecipitation - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Immunoprecipitation - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    ATM was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 at 1/40 dilution. Western blot was performed from the immunoprecipitate using Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HEK-293 whole cell lysate, 10 μg (Input).

    Lane 2: Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    ATM cleavage has been documented previously and the fragment pattern is consistent with what has been described in the literature PMID:16849690

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

    All lanes: Immunoprecipitation - Anti-ATM antibody [EPR20100] - ChIP Grade (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022)

    Predicted band size: 351 kDa

    Observed band size: 350 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    This ICC data was generated using the same anti-ATM antibody clone [EPR20100] in a different buffer formulation (cat# Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) cells labeling ATM with Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on SH-SY5Y cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling ATM with Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 at 1/800 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

  • Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533), expandable thumbnail

    Western blot - Anti-ATM antibody [EPR20100] - BSA and Azide free (ab223533)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022).

    Anti-ATM antibody [EPR20100] (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022) 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, Anti-ATM antibody [EPR20100] - ChIP Grade ab201022 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 [EPR20100] - ChIP Grade (Anti-ATM antibody [EPR20100] - ChIP Grade ab201022) 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

    Performed under reducing conditions.

    Predicted band size: 351 kDa

    Observed band size: 350 kDa

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