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74965
PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet #74965

Citations (1)
Western blot analysis of extracts from M059K cells, which express normal levels of DNA-PKcs, and M059J cells, which lack DNA-PKcs, using DNA-PKcs (E6U3A) Rabbit mAb (upper) or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Western blot analysis of extracts from M059K (DNA-PKcs positive) and M059J (DNA-PKcs negative) cells, untreated (-), or treated with neocarzinostatin (0.5 µg/ml, 1 hr; +) to induce DNA double strand breaks, using Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb (upper), total DNA-PKcs (E6U3A) Rabbit mAb #38168 (middle), or α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human colon using DNA-PKcs (E6U3A) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometriod carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded M059K cell pellet (left, positive) or M059J cell pellet (right, negative) using DNA-PKcs (E6U3A) Rabbit mAb.
Confocal immunofluorescent analysis of M059K cells (left, positive) and M059J cells (right, negative) using DNA-PKcs (E6U3A) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
Flow cytometric analysis of M059J cells (blue) and M059K cells (green) using DNA-PKcs (E6U3A) Rabbit mAb (solid lines) or a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 74965
Cat. # Size Qty. Price
74965S
1 Kit

Product Includes Quantity Reactivity MW(kDa) Isotype
DNA-PKcs (E6U3A) Rabbit mAb 38168 100 µl H 450 Rabbit IgG
Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb 68716 100 µl H 450 Rabbit IgG

Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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).

  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.

Pathways

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Limited Uses

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