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Technical Data Sheet

AM3661

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ATM (Ser-1981), phospho-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms
370 kDa

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Enlarge

Western blot of human A431 cells treated with Calyculin A (100 nM) for 30 min. Blot lanes were untreated (lanes 1, 3, & 5) or treated with lambda phosphatase (lanes 2, 4, & 6) then probed with anti-ATM (Ser-794) (lanes 1 & 2), anti-ATM (C-Terminal) (lanes 3 & 4), or anti-ATM (Ser-1981) (lanes 5 & 6).

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Enlarge

Immunocytochemical labeling of ATM phosphorylation in control (Top row) or calyculin A-treated A431 cells (Bottom row). The cells were labeled with mouse monoclonal ATM (C-terminal region) (AM3611) and ATM (Ser-1981) (AM3661). The antibodies were detected using goat anti-mouse-DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
1:200


IP
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
Ataxia telangiectasia mutated kinase (ATM) is a serine/threonine kinase that regulates cell cycle checkpoints and DNA repair. Mutations of ATM cause a spectrum of defects ranging from neurodegeneration to cancer predisposition. Activation of ATM after DNA damage involves Cdk5 mediated phosphorylation of Ser-794 followed by autophosphorylation at Ser-1891. Active ATM kinase regulates a number of proteins involved in cell cycle checkpoint control, apoptosis and DNA repair. The Cdk5–ATM pathway regulates phosphorylation and function of the ATM targets p53 and H2AX in postmitotic neurons. Other known substrates of ATM include Chk2, Chk1, CtIP, 4E-BP1, BRCA1, RPA3, SMC1, FANCD2, Rad17, Artemis, Nbs1, and the I-2 regulatory subunit of PP1. Thus, activation of Cdk5 by DNA damage may be an important initiator of ATM-dependent regulation of cell cycle checkpoints.


Background References
Shiloh, Y. (1997) Annu Rev Genet. 31:635.
Lee, J.H. & Paull, T.T. (2007) Oncogene 26:7741.
Tian, B. et al. (2009) Nat Cell Biol. 11:211.
Immunogen
Clone M366 was generated from a phospho-peptide that included amino acids surrounding Serine 1981 in human ATM. This sequence has high homology to the conserved site in rat and mouse ATM.
Buffer and Storage
Mouse monoclonal, protein A purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at -20°C. Stable for 1 year.
Specificity
This antibody detects a 370 kDa* protein corresponding to the molecular mass of ATM on SDS-PAGE immunoblots of calyculin A treated human A431 and Jurkat Cells, but is not observed in control cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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