ATM Antibody (2C1) - Azide and BSA Free

Novus Biologicals | Catalog # NB100-309

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey

Cited:

Human, Mouse, Hamster - Cricetulus (Chinese Hamster)

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Knockdown Validated, SDS-Page, Single Cell Western

Cited:

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

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 2C1

Format

Azide and BSA Free
View all ATM Primary Antibodies »

Product Specifications

Immunogen

Recombinant protein expressed in E. coli corresponding to amino acids 2577-3056.

Reactivity Notes

Use in Human reported in scientific literature (PMID:33743824).

Localization

Nucleus. Cytoplasmic vesicle.

Specificity

ATM Antibody (2C1) recognizes full-length ATM, a 350-kDa nuclear phosphoprotein.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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 (2C1) - Azide and BSA Free

Western Blot: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309] - Detection of human ATM protein using ATM antibody (2C1) [NB100-309] by western blot or immunoprecipitation. Theoretical molecular weight 351 kDa.
Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309]

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309]

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309] - Human breast carcinoma. ATM stained by ATM antibody [2C1] diluted at 1:100.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
Western Blot: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309] - Analysis of ATM in U2OS sarcoma cells using ATM antibody (2C1) [NB100-309]. Theoretical molecular weight 351 kDa. Western blot image submitted by a verifeid customer review.
Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309]

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309]

Immunohistochemistry-Paraffin: ATM Antibody (2C1) [NB100-309] - Human Kidney (formalin-fixed, paraffin-embedded) stained with ATM antibody (2C1) [NB100-309] at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.
Knockdown Validated: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (60 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody (2C1) [NB100-309] diluted at 1:500.
Knockdown Validated: ATM Antibody (2C1) [NB100-309]

Western Blot: ATM Antibody (2C1) [NB100-309]

ATM-Antibody-2C1-Western-Blot-NB100-309-img0019.jpg
Immunoprecipitation: ATM Antibody (2C1) [NB100-309]

Immunoprecipitation: ATM Antibody (2C1) [NB100-309]

Immunoprecipitation: ATM Antibody (2C1) [NB100-309] - MDA-MB-231 whole cell lysates were immunoprecipitated with 2 ug ATM Antibody (NB100-309), followed by Western blot (primary antibody: NB100-309 at 1:1000, 4C overnight. Western blot image submitted by a verified customer review.
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] - 

nb100-309_mouse-monoclonal-atm-antibody-2c1-25520231514910.jpg
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] -

Western Blot: ATM Antibody (2C1) [NB100-309] - Western blot analysis of the extent of knockdown MRE11 & ATM, & the lack of effect of Mirin activation of ATM. The extent of shRNA-mediated knockdown of MRE11 & ATM are shown for (A) clones GFP-7F1 & GFP-6D1 & (B) clones EDS-7F2 & EDS-6J8. The extent of knockdown was determined by analyzing the intensity of the ATM & MRE11 bands relative to the intensity of the loading control GAPDH bands (see Table 1). (C) The effect of Mirin on activation of ATM was determined by analysis of phosphorylation of ATM in response to ionizing radiation. Cultures treated with DMSO alone, 20 μM Mirin, or knockdown of ATM were analyzed by western blot 30 min after exposure to 10 Gy of ionizing radiation. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26209132), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] -

Western Blot: ATM Antibody (2C1) [NB100-309] - Rac1 inhibition abolishes IR-induced activation of both ATM & ATR signaling. (A) MCF-7 cells were treated with/without 20-Gy IR in the presence or absence of 100 μM NSC23766 & incubated for 1 hour at 37°C before analysis. To assess the ATM kinase activity, ATM was immunoprecipitated from cell lysate by using anti-ATM antibody (2C1) & assayed for ATM activity by using p53 recombinant protein as substrate. To measure the Chk2 activity, Chk2 was immunoprecipitated from cell lysate by using B-4 anti-Chk2 antibody & assayed for Chk2 activity by using Cdc25C recombinant protein as substrate. As controls, ATM & Chk2 protein levels in the immunoprecipitates (IP-WB) as well as in cell lysates (WB) were assessed with immunoblotting. (B) ATR & Chk1 were immunoprecipitated from cell lysates by using N-19 anti-ATR & G-4 anti-Chk1 antibody, respectively. ATR activity was assayed by using p53 recombinant protein substrate, & Chk1 activity assayed by using Cdc25C recombinant protein substrate. As controls, ATR & Chk1 protein levels in the immunoprecipitates (IP-WB), as well as in cell lysates (WB) were assessed with immunoblotting. (C) MCF-7 cells were exposed to IR at the indicated doses in the presence or absence of NSC23766, incubated for 1 hour, & assessed for Chk1 & Chk2 activities. *Kinase assay does not contain Cdc25C substrate. (D) T47D & ZR-75-1 cells were exposed to 10-Gy IR in the presence or absence of 100 μM NSC23766, incubated for 1 hour, & analyzed for Chk1 & Chk2 activities. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] -

Western Blot: ATM Antibody (2C1) [NB100-309] - Inhibition of Rac1 by N17Rac1 mutant or Rac1 siRNA diminishes IR-induced G2/M checkpoint activation. (A) MCF-7 cells were infected with Ad.N17Rac1 or Ad.Control for 24 hours & exposed to 15-Gy IR. Left panel: the cells were analyzed for DNA content 24 hours after IR. The result depicts the percentage of cells with 4N-DNA content & is shown as mean ± SD of quadruplicate samples. *P < 0.001 (n = 4), significant difference from the irradiated Ad.Control-infected cells. Right panel: Inset: at 15 minutes after IR, the infected cells were analyzed for Rac1 activities (Rac1-GTP) & protein levels (total Rac1). Bar graph: mitotic cells in the cell samples were analyzed 2 hours after IR. The result depicts the percentage of mitotic cells & is shown as mean ± SD of triplicate samples. **P = 0.002 (n = 3), significant difference from the irradiated Ad.Control-infected cells. (B) Upper panel: MCF-7 cells transfected with Rac1 siRNA (Rac1) or control siRNA (Control) were incubated for the indicated times & analyzed for protein levels of Rac1 & Actin. Lower panel: After 2-day incubation, the siRNA-transfected cells were exposed to IR, incubated for 24 hours, & assessed for DNA content. Results depict the percentage of cells with 4N-DNA content & represent the mean ± SD of three separate experiments in duplicate samples. *P < 0.001 (n = 6), significant difference from the irradiated Control-siRNA transfected cells. (C) After 2-day incubation, siRNA-transfected cells were treated with/without 20-Gy IR, incubated for 1 hour, & analyzed for ATM, ATR, Chk1, & Chk2 activities. Image collected & cropped by CiteAb from the following publication (http://breast-cancer-research.biomedcentral.com/articles/10.1186/bcr3164), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] -

Western Blot: ATM Antibody (2C1) [NB100-309] - HeLa whole cell extract and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:500. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
ATM Antibody (2C1)

Western Blot: ATM Antibody (2C1) [NB100-309] -

Western Blot: ATM Antibody (2C1) [NB100-309] - Whole cell extract (30 ug) was separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody [2C1] (NB100-309) diluted at 1:1000.
ATM Antibody (2C1) - Azide and BSA Free

Immunohistochemistry: ATM Antibody (2C1) [NB100-309] -

Human Testis (formalin-fixed, paraffin-embedded) stained with ATM antibody at 5 ug/ml followed by biotinylated anti-mouse IgG secondary antibody, alkaline phosphatase-streptavidin and chromogen.

Applications for ATM Antibody (2C1) - Azide and BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

ELISA

1:100 - 1:2000

Immunocytochemistry/ Immunofluorescence

1:10 - 1:500

Immunohistochemistry

5 ug/mL

Immunohistochemistry-Paraffin

5 ug/mL

Immunoprecipitation

1 - 10 ug/mL

Single Cell Western

100 ug/mL

Western Blot

1:500 - 1:3000
Application Notes
Use in SDS-PAGE reported in scientific literature (PMID:34210973). Use in IHC-P reported in scientific literature (PMID: 25895060). Use in FLOW reported in scientific literature (PMID: 15197179). ATM antibody is validated for WB, IP from a verified customer reviews.

Reviewed Applications

Read 3 reviews rated 4.7 using NB100-309 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. 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); 11920 (Mouse); 300711 (Rat)

Gene Symbol

ATM

UniProt

Q13315

Additional ATM Products

Product Documents for ATM Antibody (2C1) - Azide and BSA Free

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Product Specific Notices for ATM Antibody (2C1) - Azide and 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 (2C1) - Azide and BSA Free

Customer Reviews for ATM Antibody (2C1) - Azide and BSA Free (3)

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • ATM Antibody (2C1)
    Name: Yongkang Yang
    Application: Immunoprecipitation
    Sample Tested: MDA-MB-231
    Species: Human
    Verified Customer | Posted 10/28/2020
    Western Blot: MDA-MB-231 whole cell lysates were IP with 2μg ATM Antibody (NB100-309), followed by Western blot (primary antibody: NB100-309， 1:1000, 4℃, overnight).
    ATM Antibody (2C1) - Azide and BSA Free NB100-309
  • Name: Anonymous
    Application: Western Blot
    Sample Tested:
    Species: Human
    Verified Customer | Posted 04/28/2015
    ATM antibody (NB100-309)
    ATM Antibody (2C1) - Azide and BSA Free NB100-309
  • Name: Takeshi Shi
    Application: Western Blot
    Sample Tested: U2OS whole cell lysate
    Species: Human
    Verified Customer | Posted 11/02/2014

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Protocols

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FAQs for ATM Antibody (2C1) - Azide and BSA Free

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

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

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