Revision 5

#38168

Store at -20C

DNA-PKcs (E6U3A) Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IHC-P, IF-IC, FC-FP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
450

Source/Isotype:
Rabbit IgG

UniProt ID:
#P78527

Entrez-Gene Id:
5591

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #58268.

Specificity/Sensitivity

DNA-PKcs (E6U3A) Rabbit mAb recognizes endogenous levels of total DNA-PKcs protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro608 of human DNA-PKcs protein.

Background

DNA-dependent protein kinase (DNA-PK) is an important factor in the repair of double-stranded breaks in DNA. Cells lacking DNA-PK or in which DNA-PK is inhibited fail to show proper nonhomologous end-joining (NHEJ) (1-7). DNA-PK is composed of two DNA-binding subunits (Ku70 and Ku86) and one 450 kDa catalytic subunit (DNA-PKcs) (8). It is thought that a heterodimer of Ku70 and Ku86 binds to double-stranded DNA broken ends before DNA-PKcs binds and is activated (1,9). Activated DNA-PKcs is a serine/threonine kinase that has been shown to phosphorylate a number of proteins in vitro, including p53, transcription factors, RNA polymerase, and Ku70/Ku86 (10,11). DNA-PKcs autophosphorylation at multiple sites, including Thr2609 and Ser2056, results in an inactivation of DNA-PK kinase activity and NHEJ ability (12,13). It has been demonstrated, however, that DNA-PK preferentially phosphorylates substrates before it autophosphorylates, suggesting that DNA-PK autophosphorylation may play a role in disassembly of the DNA repair machinery (14,15). Autophosphorylation at Thr2609 has also been shown to be required for DNA-PK-mediated double-strand break repair, and phosphorylated DNA-PK co-localizes with H2A.X and 53BP1 at sites of DNA damage (16). Phosphorylation at Ser2056 occurs in response to double-stranded DNA breaks and ATM activation (17).

Background References

  1. Gottlieb, T.M. and Jackson, S.P. (1993) Cell 72, 131-42.
  2. Hartley, K.O. et al. (1995) Cell 82, 849-56.
  3. Rosenzweig, K.E. et al. (1997) Clin Cancer Res 3, 1149-56.
  4. Jackson, S.P. and Jeggo, P.A. (1995) Trends Biochem Sci 20, 412-5.
  5. Roth, D.B. et al. (1995) Curr Biol 5, 496-9.
  6. Baumann, P. and West, S.C. (1998) Proc Natl Acad Sci U S A 95, 14066-70.
  7. Chen, S. et al. (2001) J Biol Chem 276, 24323-30.
  8. Jeggo, P.A. (1997) Mutat Res 384, 1-14.
  9. Suwa, A. et al. (1994) Proc Natl Acad Sci U S A 91, 6904-8.
  10. Anderson, C.W. and Lees-Miller, S.P. (1992) Crit Rev Eukaryot Gene Expr 2, 283-314.
  11. Kuhn, A. et al. (1995) Genes Dev 9, 193-203.
  12. Chan, D.W. and Lees-Miller, S.P. (1996) J Biol Chem 271, 8936-41.
  13. Douglas, P. et al. (2002) Biochem. J. 368, 243-51.
  14. Lees-Miller, S.P. et al. (1992) Mol Cell Biol 12, 5041-9.
  15. Jackson, S.P. et al. (1990) Cell 63, 155-65.
  16. Chan, D.W. et al. (2002) Genes Dev 16, 2333-8.
  17. Yajima, H. et al. (2009) J Mol Biol 385, 800-10.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.