ATM Antibody (5C2) - Azide and BSA Free

Novus Biologicals | Catalog # NB100-220

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey

Cited:

Human, Mouse, Avian - Chicken

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 5C2

Format

Azide and BSA Free
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Product Specifications

Immunogen

Recombinant protein expressed in E. coli corresponding to amino acids 980-1512.

Reactivity Notes

Human and Mouse reactivity reported in multiple pieces of scientific literature. Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Mouse-On-Mouse blocking reagent may be needed for IHC and ICC experiments to reduce high background signal. You can find these reagents under catalog numbers PK-2200-NB and MP-2400-NB. Please contact Technical Support if you have any questions.

Localization

Nuclear

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 (5C2) - Azide and BSA Free

Western Blot: ATM Antibody (5C2) [NB100-220]

Western Blot: ATM Antibody (5C2) [NB100-220]

Western Blot: ATM Antibody (5C2) [NB100-220] - HeLa whole cell and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with ATM antibody (5C2) [NB100-220] diluted at 1:1000. The HRP-conjugated anti-mouse IgG antibody [NBP2-19382] was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced. Observed molecular weight ~320 kDa. Theoretical molecular weight 351 kDa.
Immunocytochemistry/ Immunofluorescence: ATM Antibody (5C2) [NB100-220]

Immunocytochemistry/ Immunofluorescence: ATM Antibody (5C2) [NB100-220]

Immunocytochemistry/Immunofluorescence: ATM Antibody (5C2) [NB100-220] - HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: ATM protein stained by ATM antibody (5C2) [NB100-220] diluted at 1:200. Blue: Hoechst 33342 staining. Scale bar = 10 um.
Western Blot: ATM Antibody (5C2) [NB100-220]

Western Blot: ATM Antibody (5C2) [NB100-220]

ATM-Antibody-5C2-Western-Blot-NB100-220-img0008.jpg
ATM Antibody (5C2)

Western Blot: ATM Antibody (5C2) [NB100-220] -

Western Blot: ATM Antibody (5C2) [NB100-220] - Absence of Atm does not relieve the selective pressure to inactivate p53 during chemical carcinogenesis.A. Representative images of 3MC-fibrosarcomas immunostained for Arf, p53 & p21 (see also Table 1). The upper & middle rows are representative of the large majority of fibrosarcomas developed in wild-type (upper) & Atm-null (middle) mice, which are consistent with a mutant p53 (i.e. strongly positive for p53 & negative for p21). The lower row is representative of the fibrosarcomas developed in Arf-null mice, which are consistent with a functional p53 (i.e. very weakly positive for p53 & positive for p21). B. Examples of cancer cell lines established from 3MC-fibrosarcomas (each line derives from an independent fibrosarcoma). The genotype of the mice where the 3MC-fibrosarcomas were generated is indicated, as well as, the status of p53 as determined by a nutlin-sensitivity assay (see Supplementary Figures S5 & S6). Cell lines were exposed to 10 Gy & protein extracts were obtained 1h & 24h after irradiation. The levels of the indicated proteins were determined by immunoblotting using beta -actin as loading control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/19421407), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody (5C2)

Immunohistochemistry: ATM Antibody (5C2) [NB100-220] -

Immunohistochemistry: ATM Antibody (5C2) [NB100-220] - Limited role of Atm in p53-mediated tumor suppression & DNA damage response in chemically-induced fibrosarcomas.A. Mice of the indicated genotypes, wt (n = 15), p53-super (n = 16), Atm-null (n = 9), p53-super/Atm-null (n = 13) & Arf-null (n = 12), were injected intramuscularly with 3-methyl-cholanthrene (3MC) & tumour development was monitored. Kaplan-Meier tumour-free curves were obtained & statistical significant differences (logrank test) were found for wt vs. p53-super (p<0.005), Atm-null vs. p53-super/Atm-null (p<0.001) & Arf-null vs. wt (p<0.0001). No significant differences were found for wt vs. Atm-null (p = 0.11), or p53-super vs. p53-super/Atm-null (p = 0.18). B. Quantification of the persistent DNA damage response ( gamma H2AX), proliferation (Ki67), & apoptosis (activated caspase-3) in 3MC-fibrosarcomas generated in mice of the indicated genotypes. Quantifications are relative to the 3MC-fibrosarcomas in wild-type mice. Values correspond to the average & standard deviation (n = 5 per genotype). Examples of the immunostainings are shown in the panels at the right. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/19421407), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
ATM Antibody (5C2)

Immunocytochemistry/ Immunofluorescence: ATM Antibody (5C2) [NB100-220] -

Immunocytochemistry/ Immunofluorescence: ATM Antibody (5C2) [NB100-220] - BAG3P209L aggregation leads to co-aggregation of proteasomal substrates. a Immunofluorescence pictures of HeLa cells expressing FLAG-BAG3WT or FLAG-BAG3P209L using antibodies against BAG3 (green) or ubiquitin (red). Scale bar = 5 μm. b Suppression of GFP-HttQ74 aggregation of cells expressing a control, FLAG-BAG3WT or FLAG-BAG3P209L. Western blot against indicated antibodies is shown. c Quantification of GFP-HttQ74 aggregation of experiments similar to b. Relative percentage of SDS-insoluble protein levels are shown. Data represents the mean & standard deviation of three independent experiments. d Immunofluorescence pictures of HeLa cells expressing FLAG-BAG3WT or FLAG-BAG3P209L using antibodies against BAG3 (green) or ubiquitin (red). Left column BAG3, middle column ubiquitin & right column is the merge of BAG3 (green), ubiquitin (red), & DAPI (blue). Scale bar = 5 μm. e Fractionation of HEK293 cells expressing HSPB8, a control or BAG3WT or BAG3P209L, together with either Ub-R-GFP or GFP-ODC (ornithine decarboxylase). Western blot against GFP, FLAG (BAG3), Myc (HSPB8), & tubulin are shown. f Fractionation of HEK293 cells expressing HSPB8 & indicated BAG3 variants. Western blot using ubiquitin (FK2) & tubulin antibodies are shown. The same samples as in Fig. 5e have been used, loading control is therefore the same. Source data are provided as a Source data file Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30559338), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for ATM Antibody (5C2) - Azide and BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Western Blot

1:500-1:3000
Application Notes
Use in ICC/IF reported in (PMID: 26447695). Use in WB reported in (PMID: 22869595). IHC, IP-Assay dependent.

Formulation, Preparation, and Storage

Purification

Protein G 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

Additional ATM Products

Product Documents for ATM Antibody (5C2) - Azide and BSA Free

Certificate of Analysis

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

Product Specific Notices for ATM Antibody (5C2) - 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

Citations for ATM Antibody (5C2) - Azide and BSA Free

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Protocols

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

FAQs for ATM Antibody (5C2) - 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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