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Technical Data Sheet

AP3631

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ATM (Ser-794), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
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 calyculin A-treated A431 cells. The cells were labeled with rabbit polyclonal anti-ATM (Ser-794) (AP3631) antibody in the absence (Left) or presence (Right) of blocking peptide (AX3635). The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
1:200


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
Phospho-ATM (Ser-794) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding Ser-794 in human ATM. This sequence is well conserved in rat and mouse ATM.
Buffer and Storage
Rabbit polyclonal, affinity-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 was affinity purified using phospho-ATM (Ser-794) peptide (without carrier). The antibody detects a 370 kDa* band corresponding to ATM on SDS-PAGE immunoblots of calyculin A treated Jurkat, A431, HeLa, and rat PC12 cells. This reactivity is removed after lambda phosphatase treatment.

*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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