Invitrogen

Phospho-ATM (Ser1981) Monoclonal Antibody (10H11.E12), eFluor™ 660, eBioscience™

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Datasheet
Protocols
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Datasheet
Protocols
Questions & Answers

Product Details

50-9046-42

Applications
Tested Dilution
Publications

Immunohistochemistry (IHC)

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Flow Cytometry (Flow)

5 µL (0.25 µg)/test
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Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Human

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), eFluor™ 660, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

10H11.E12

Conjugate

eFluor™ 660 eFluor™ 660 eFluor™ 660

Excitation/Emission Max

651/668 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2574313

Product Specific Information

Description: This 10H11.E12 monoclonal antibody recognizes the human, mouse, and rat ataxia-telangiectasia (ATM) protein when phosphorylated on serine 1981. ATM belongs to the family of PI3 kinases and functions by coordinating cell cycle arrest and initiation of DNA repair through the phosphorylation of a multitude of substrates in response to DNA damage and oxidative stress. ATM exists as dimers or multimers in its inactive state but in response to DNA breaks, becomes activated through formation of monomers and autophosphorylation. Activated ATM is recruited to the site of DNA double strand breaks where it halts the cell cycle and initiates DNA repair through the phosphorylation of downstream DNA damage response pathway members. Loss of functional activity of ATM results in ataxia-telangiectasia (AT), a disease characterized by early onset neurodegeneration and predisposition to cancer. AT patients are immunodeficient, radiosensitive, and have an increased risk of certain cancer types such as lymphoma and leukemia.

Applications Reported:This 10H11.E12 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 10H11.E12 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochrome.

Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the BestProtocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the BestProtocols® Section under the Resources tab online.

Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Ataxia-telangiectasia Mutated (ATM) is a protein that belongs to the PI3/PI4 kinase family. Ataxia-telangiectasia is a rare autosomal recessive disorder characterized by progressive neurologic degeneration, immunologic deficiency, and an increased risk of lymphoid cancer. The ATM gene codes for a protein belonging to the phosphoinositide 3-kinase (PI3K) superfamily. ATM phosphorylates proteins instead of lipid and has many downstream targets that act as cell-cycle regulators including: P53, Mdm2, BRCA1, and SMC1. The ATM protein is responsible for repairing double-stranded DNA breaks that occur because of ionizing radiation and other mutagens. The ATM’s C-terminal region has extensive homology to the catalytic domains of phosphatidylinositol 3-kinases (PI3 kinases). Studies have shown that ATM becomes autophosphorylated and upregulated by exposure to ionizing radiation. AT cells are hypersensitive to ionizing radiation, impaired in mediating the inhibition of DNA synthesis and display delays in p53 induction. Further, DNA damage caused by ionizing irradiation activates ATM-kinase, leading to a cascade of kinase reactions that regulate cell cycle, apoptosis, and DNA damage repair. Studies have linked ATM to apoptosis along with Nbs1 and Chk2 in the E2F1 pathway.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: A-T mutated; A-T mutated homolog; AT mutated; AT mutated protein; Ataxia t; Ataxia telangiectasia gene mutated in human beings; Ataxia telangiectasia mutated; Ataxia telangiectasia mutated homolog; DKFZp781A0353; EC 2.7.1.37; MGC74674; Serine-protein kinase ATM; TEL1, telomere maintenance 1, homolog

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Gene Aliases: AI256621; AT1; ATA; ATC; ATD; ATDC; ATE; ATM; C030026E19Rik; TEL1; TELO1

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UniProt ID: (Human) Q13315, (Mouse) Q62388

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Entrez Gene ID: (Human) 472, (Mouse) 11920, (Rat) 300711

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Function(s)
DNA binding protein serine/threonine kinase activity DNA-dependent protein kinase activity protein binding ATP binding 1-phosphatidylinositol-3-kinase activity protein complex binding protein dimerization activity protein N-terminus binding nucleotide binding protein kinase activity kinase activity transferase activity transferase activity, transferring phosphorus-containing groups phosphotransferase activity, alcohol group as acceptor histone serine kinase activity
Process(es)
DNA damage checkpoint telomere maintenance double-strand break repair via homologous recombination response to hypoxia somitogenesis pre-B cell allelic exclusion DNA repair protein phosphorylation cellular response to DNA damage stimulus DNA damage induced protein phosphorylation cell cycle cell cycle arrest mitotic spindle assembly checkpoint female gamete generation brain development heart development determination of adult lifespan intrinsic apoptotic signaling pathway in response to DNA damage response to ionizing radiation phosphorylation histone phosphorylation peptidyl-serine phosphorylation negative regulation of B cell proliferation V(D)J recombination histone-serine phosphorylation phosphatidylinositol-3-phosphate biosynthetic process lipoprotein catabolic process positive regulation of apoptotic process positive regulation of DNA damage response, signal transduction by p53 class mediator positive regulation of neuron apoptotic process protein autophosphorylation phosphatidylinositol phosphorylation oocyte development neuron apoptotic process regulation of cell cycle histone mRNA catabolic process cellular response to gamma radiation signal transduction involved in mitotic G2 DNA damage checkpoint replicative senescence DNA double-strand break processing DNA synthesis involved in DNA repair strand displacement DNA replication double-strand break repair via nonhomologous end joining DNA damage response, signal transduction by p53 class mediator resulting in cell cycle arrest telomere maintenance via telomerase reciprocal meiotic recombination signal transduction regulation of autophagy positive regulation of telomere maintenance via telomerase positive regulation of histone phosphorylation peptidyl-serine autophosphorylation regulation of apoptotic process meiotic telomere clustering regulation of telomerase activity cellular response to X-ray cellular response to nitrosative stress establishment of RNA localization to telomere establishment of macromolecular complex localization to telomere regulation of cellular response to heat regulation of signal transduction by p53 class mediator negative regulation of TORC1 signaling negative regulation of telomere capping positive regulation of telomere maintenance via telomere lengthening positive regulation of telomerase catalytic core complex assembly apoptotic process
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