Cell Signaling Technology

Product Pathways - DNA Damage

Ku80 Antibody #2753

Applications Reactivity MW (kDa) Source
W IP IHC-P IF-IC F H Mk (M) (R) 86 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Ku80 antibody detects endogenous levels of total Ku80 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to the carboxy terminus of human Ku80. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa, A673, A549 and COS cells, using Ku80 antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Ku80 Antibody preincubated with control peptide (left) or antigen-specific peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistsochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localiztion, using Ku80 Antibody.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, using Ku80 Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-F

IF-F

DAPI staining (left) and immunofluorescent staining (right) of paraformaldehyde-fixed HeLa cells, using Ku80 Antibody.

Background

Ku is a heterodimeric protein composed of two subunits (Ku70 and Ku80) originally identified as autoantigens associated with several autoimmune diseases including scleroderma, polymyositis and systemic lupus erythematosus (1). Ku is an abundant, ubiquitously expressed nuclear protein that binds to and stabilizes the ends of DNA at telomeres or double-stranded DNA breaks (2-5). The Ku70/Ku80 heterodimer has ATP-dependent DNA helicase activity and functions as the DNA-binding regulatory component of DNA-dependent protein kinase (DNA-PK) (6-8). The assembly of DNA-PK complex at DNA ends is required for nonhomologous end-joining (NHEJ), one mechanism involved in double-stranded DNA break repair and V(D)J recombination (8). DNA-PK has been shown to phosphorylate many proteins, including p53, serum response factor, c-Jun, c-Fos, c-Myc, Oct-1, Sp-1 and RNA polymerase II (1,8). The combined activities of Ku70/Ku80 and DNA-PK implicate Ku in many cellular functions, including cell-cycle regulation, DNA replication and repair, telomere maintenance, recombination and transcriptional activation.

  1. Tuteja, R. and Tuteja, N. (2000) Crit. Rev. Biochem. Mol. Biol. 35, 1-33.
  2. Blier, P.R. et al. (1993) J. Biol. Chem. 268, 7594-7601.
  3. Jin, S. and Weaver, D.T. (1997) EMBO J. 16, 6874-6885.
  4. Boulton, S.J. and Jackson, S.P. (1998) EMBO J. 17, 1819-1828.
  5. Gravel, S. et al. (1998) Science 280, 741-744.
  6. Cao, Q.P. et al. (1994) Biochemistry 33, 8548-8557.
  7. Lees-Miller, S.P. et al. (1990) Mol. Cell Biol. 10, 6472-6481.
  8. Collis, S.J. et al. (2005) Oncogene 24, 949-961.

Application References

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