Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Canine, Kangaroo

Cited:

Human, Mouse, Canine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoblotting, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot, Immunoblotting, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all ATM Primary Antibodies »

Product Specifications

Immunogen

ATM Antibody was made to a fragment of the human ATM protein corresponding to the C-terminus (within the last third of the protein sequence). [Uniprot: Q13315]

Reactivity Notes

Canine reactivity reported in scientific literature (PMID: 31648115).

Localization

Nuclear

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

351 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for ATM Antibody - BSA Free

Western Blot: ATM Antibody [NB100-104]

Western Blot: ATM Antibody [NB100-104]

Western Blot: ATM Antibody [NB100-104] - Detection of ATM in HeLa nuclear extract using ATM antibody [NB100-104]. Theoretical molecular weight 351 kDa.
Immunohistochemistry-Paraffin: ATM Antibody [NB100-104]

Immunohistochemistry-Paraffin: ATM Antibody [NB100-104]

Immunohistochemistry-Paraffin: ATM Antibody [NB100-104] - Staining of human tonsil, germinal center and mantle zone with ATM Antibody [NB100-104].
Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104]

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104]

ATM-Antibody-Immunocytochemistry-Immunofluorescence-NB100-104-img0018.jpg
Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104] - An intracellular stain was performed on HeLa cells with Dylight 550-conjugated ATM Antibody [NB100-104R] (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.
Western Blot: ATM Antibody [NB100-104]

Western Blot: ATM Antibody [NB100-104]

ATM-Antibody-Western-Blot-NB100-104-img0017.jpg
Western Blot: ATM Antibody [NB100-104]

Western Blot: ATM Antibody [NB100-104]

Western Blot: ATM Antibody [NB100-104] - Detection of ATM in Raji whole cell lysate using [NB100-104]. Observed molecular weight ~300 kDa. Theoretical molecular weight 351 kDa.
Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104]

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104]

Immunocytochemistry/Immunofluorescence: ATM Antibody [NB100-104] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.05% Triton X-100. Then cells were incubated with [NB100-104] at a 1:100 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104] - Analysis of PE conjugate of ATM Antibody [NB100-104PE]. An intracellular stain was performed on HeLa cells with ATM antibody [NB100-104PE] (blue) and a matched isotype control [NBP2-24893PE] (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin.
Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104]

Flow Cytometry: ATM Antibody [NB100-104] - An intracellular stain was performed on HepG2 cells with PE-conjugated ATM antibody [NB100-104PE] (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Phycoerythrin.
ATM Antibody

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104] -

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104] - ATM is activated by spermidine in GM00637 cells.GM00637 cells with or without KU55933 pretreatment (a,b), & GM05849 cells (c) were exposed to 50 μM spermidine or CCCP, followed by immunofluorescence analyses of total & p-ATM on Ser-1981. The scale bar is 10 μm. Ratios of cells expressing p-ATM Ser-1981 to cells expressing total ATM were presented (d). 20–45 cells/condition from three experiments were collected. Values are mean ± SD, *p < 0.05 vs. control. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep24700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104] -

Immunocytochemistry/ Immunofluorescence: ATM Antibody [NB100-104] - ATM is activated by spermidine in GM00637 cells.GM00637 cells with or without KU55933 pretreatment (a,b), & GM05849 cells (c) were exposed to 50 μM spermidine or CCCP, followed by immunofluorescence analyses of total & p-ATM on Ser-1981. The scale bar is 10 μm. Ratios of cells expressing p-ATM Ser-1981 to cells expressing total ATM were presented (d). 20–45 cells/condition from three experiments were collected. Values are mean ± SD, *p < 0.05 vs. control. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/srep24700), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody

Western Blot: ATM Antibody [NB100-104] -

Western Blot: ATM Antibody [NB100-104] - Depletion of SMARCAL1 induces DNA damage & checkpoint activation in iSML1 iPSCs. (A,B) The iSML1 iPSCs were cultured for 7 & 14 days in the presence of doxycycline (DOX) to induce SMARCAL1 downregulation & then immunostained. The graphs (top) show quantification of the number of gamma -H2AX-positive cells (A) or ATM-pSer1981-positive cells (B). Representative images from triplicate experiments are shown (bottom). (C) Immunoblot detection of the indicated DDR proteins in iSML1 iPSCs after 14 days of continuous treatment with DOX. Lamin B1 was used as the loading control. Data are mean±s.e.m. from three independent experiments. *P≤0.05, **P≤0.01, ***P≤0.001 (two-way ANOVA test). ns, not significant. Scale bars: 10 µm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31515241), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody

Western Blot: ATM Antibody [NB100-104] -

Western Blot: ATM Antibody [NB100-104] - Analysis of double strand break repair in MCF-7 monolayer & mammosphere populations. (a) Cells were exposed to 4 Gy 60Co gamma -radiation & the relative degree of double-strand breakage (DSB) was determined by the comet assay under neutral conditions immediately after exposure & at the times indicated after exposure. (b) The 'comets' (n of about 100) were categorized according to the NIH LISTSERV (Comet Assay Interest Group web site) in which type 1 comets display the least DNA damage & type 5 the most. The error bars represent the mean ± standard error of the mean in both panels. The comets of the unirradiated cells are labeled Cont. (c) Expression of proteins involved in the NHEJ pathway of DSB repair in response to increasing doses of ionizing radiation. Lysates were prepared from unirradiated cells & from cells harvested one hour after exposure to 1 or 10-Gy 60Co gamma -radiation & analyzed by immunoblotting with antibodies against several DSB repair proteins. Phospho-ATM & phospho-DNA-PKcs refer to phosphorylation of these proteins at Ser1981 & Ser2056, respectively. Actin served as a loading control. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr2583), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for ATM Antibody - BSA Free

Application
Recommended Usage

Immunoblotting

reported in scientific literature (PMID 28512243)

Immunocytochemistry/ Immunofluorescence

1 - 2 ug/ml

Immunohistochemistry

1:100 - 1:200

Immunohistochemistry-Paraffin

1:100 - 1:200

Immunoprecipitation

1:10-1:500

Western Blot

1:500-1:1000
Application Notes
In Western blot, it detects a band at ~350 kDa, representing ATM.

Reviewed Applications

Read 2 reviews rated 4.5 using NB100-104 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: ATM

ATM (ataxia telangiectasia mutated kinase) is the master regulator of the DNA double-strand break (DSB) repair pathway. This ubiquitously expressed serine/threonine protein kinase belongs to the PI3K-like family of proteins and responds to DSBs caused by oxidative and other genotoxic stresses (1). In addition to regulating the DNA damage response, ATM participates in vesicle and protein transport, T-cell development, gonads/neurological function, pre-B cell allelic exclusion, cell cycle control, and acts as a tumor suppressor (2,3). Defects in ATM are associated with ataxia telangiectasia (AT), T-cell acute lymphoblastic leukemia (TALL), T-prolymphocytic leukemia (TPLL), and B-cell non-Hodgkin lymphomas (BNHL) including mantle cell lymphoma (MCL) and B-cell chronic lymphocytic leukemia (BCLL) (4).

The theoretical molecular weight of ATM is 350 kDa and it has 3 main domains: a FAT (focal adhesion targeting) domain (aa 1960-2566), a PI-3/PI-4 kinase catalytic domain (aa 2712-2962), and a C-terminal FAT domain (aa 3024-3056). ATM exists as a dimer or tetramer in its inactive state. Upon sensing DNA damage, the MRE11-RAD50-NBS1 (MRN) complex recruits ATM. The intricate process of ATM activation involves acetylation by KAT5/TIP60, autophosphorylation at Ser-1981, and dissociation into catalytically active monomers (5). Following activation, ATM phosphorylates multiple substrates such as p53/TP53 and Chk2 involved in DNA repair, checkpoint signaling, and the apoptosis pathway.

References

1. Paull TT. (2015) Mechanisms of ATM Activation. Annu Rev Biochem. 84:711-38. PMID: 25580527

2. Chaudhary MW and Al-Baradie RS. (2014) Ataxia-telangiectasia: future prospects. Appl Clin Genet. 7:159-167. PMID: 25258552

3. Stagni V, Cirotti C, and Barila D. (2018) Ataxia-Telangiectasia Mutated Kinase in the Control of Oxidative Stress, Mitochondria, and Autophagy in Cancer: A Maestro With a Large Orchestra. Front Oncol. 8:73. PMID: 29616191

4. Gumy-Pause F, Wacker P, and Sappino AP. (2004) ATM gene and lymphoid malignancies. Leukemia. 18(2):238-42. PMID: 14628072

5. Adamowicz M. (2018) Breaking up with ATM. J Immunol Sci. 2(1):26-31. PMID: 29652413

Long Name

Ataxia Telangiectasia-mutated

Alternate Names

TEL1, TELO1, TPLL

Entrez Gene IDs

472 (Human)

Gene Symbol

ATM

UniProt

Q13315

Additional ATM Products

Product Documents for ATM Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for ATM Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

p53 Pathway

Citations for ATM Antibody - BSA Free

Customer Reviews for ATM Antibody - BSA Free (2)

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Customer Images

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  • Name: Barbara Alcaraz Silva
    Application: Immunofluorescence
    Sample Tested: cell
    Species: Other
    Verified Customer | Posted 10/08/2014
  • Name: Christie Brown
    Application: Western Blot
    Sample Tested: Lymphoblastoid (LCL) nuclear lysate
    Species: Human
    Verified Customer | Posted 08/15/2013
    ATM Antibody - BSA Free NB100-104

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Protocols

View specific protocols for ATM Antibody - BSA Free (NB100-104):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 4% paraformaldehyde to the dish and fix at room temperature for 10 minutes.
2. Remove the paraformaldehyde and wash the cells in PBS.
3. Permeabilize the cells with 0.1% Triton X100 or other suitable detergent for 2 min.
4. Remove the permeabilization buffer and wash three times for 5 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 5 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 5 minutes each.
10. Counter stain DNA with DAPI if required.

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for ATM Antibody - BSA Free

Showing  1 - 4 of 4 FAQs Showing All

  • Q: What is the expiration date of this product?

    A: We guarantee the stability of this product for 1 year from the date of receipt.

  • Q: What is the isotype of this antibody?

    A: The isotype for this ATM Antibody is IgG.

  • Q: What is the theoretical molecular weight for your ATM antibodies?

    A: The theoretical molecular weight for our ATM antibodies is 351 kDa.

  • Q: Which applications can I use this antibody in?

    A: This antibody has been cited in multiple publications for use in ELISA, flow cytometry, immunoblotting, ICC/IF, immunohistochemistry, immunoprecipitation, and western blot. This antibody has also been tested in IHC-Paraffin; therefore all of these applications fall under our 100% guarantee. If you would like to test this antibody in an application or species not listed, please check out our Innovator's Reward Program.

  • Q: What is the expiration date of this product?

    A: We guarantee the stability of this product for 1 year from the date of receipt.

  • Q: What is the isotype of this antibody?

    A: The isotype for this ATM Antibody is IgG.

  • Q: What is the theoretical molecular weight for your ATM antibodies?

    A: The theoretical molecular weight for our ATM antibodies is 351 kDa.

  • Q: Which applications can I use this antibody in?

    A: This antibody has been cited in multiple publications for use in ELISA, flow cytometry, immunoblotting, ICC/IF, immunohistochemistry, immunoprecipitation, and western blot. This antibody has also been tested in IHC-Paraffin; therefore all of these applications fall under our 100% guarantee. If you would like to test this antibody in an application or species not listed, please check out our Innovator's Reward Program.

  • Q: What is the expiration date of this product?

    A: We guarantee the stability of this product for 1 year from the date of receipt.

  • Q: What is the isotype of this antibody?

    A: The isotype for this ATM Antibody is IgG.

  • Q: What is the theoretical molecular weight for your ATM antibodies?

    A: The theoretical molecular weight for our ATM antibodies is 351 kDa.

  • Q: Which applications can I use this antibody in?

    A: This antibody has been cited in multiple publications for use in ELISA, flow cytometry, immunoblotting, ICC/IF, immunohistochemistry, immunoprecipitation, and western blot. This antibody has also been tested in IHC-Paraffin; therefore all of these applications fall under our 100% guarantee. If you would like to test this antibody in an application or species not listed, please check out our Innovator's Reward Program.

  • Q: What is the expiration date of this product?

    A: We guarantee the stability of this product for 1 year from the date of receipt.

  • Q: What is the isotype of this antibody?

    A: The isotype for this ATM Antibody is IgG.

  • Q: What is the theoretical molecular weight for your ATM antibodies?

    A: The theoretical molecular weight for our ATM antibodies is 351 kDa.

  • Q: Which applications can I use this antibody in?

    A: This antibody has been cited in multiple publications for use in ELISA, flow cytometry, immunoblotting, ICC/IF, immunohistochemistry, immunoprecipitation, and western blot. This antibody has also been tested in IHC-Paraffin; therefore all of these applications fall under our 100% guarantee. If you would like to test this antibody in an application or species not listed, please check out our Innovator's Reward Program.

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