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

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

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PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 1

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.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 2

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).

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 3

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).

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 4

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 5

Immunohistochemical analysis of paraffin-embedded normal human colon using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 6

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 7

Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 8

Immunohistochemical analysis of paraffin-embedded human endometriod carcinoma using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 9

Immunohistochemical analysis of paraffin-embedded M059K cell pellet (left, positive) or M059J cell pellet (right, negative) using DNA-PKcs (E6U3A) Rabbit mAb.

PhosphoPlus® DNA-PKcs (Ser2056) Antibody Duet: Image 10

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).

To Purchase # 74965S
Product # Size Price
74965S
1 Kit $ 514

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 & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus is a trademark of Cell Signaling Technology, Inc.