Revision 1

#2753Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-P, IF-IC, FC-FP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

86

SOURCE:

Rabbit

UniProt ID:

#P13010

Entrez-Gene Id:

7520

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:25
Immunohistochemistry (Paraffin) 1:150 - 1:600
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:100

Storage

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

Specificity / Sensitivity

Ku80 antibody detects endogenous levels of total Ku80 protein.

Species Reactivity:

Human, Monkey

Species predicted to react based on 100% sequence homology

Mouse, Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of mouse Ku80. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Ku is a heterodimeric protein composed of two subunits (Ku70 and Ku80) originally identified by researchers 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 the 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.

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

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

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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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 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 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.

Revision 1
#2753

Ku80 Antibody

Western Blotting Image 1: Ku80 Antibody Expand Image
Western blot analysis of lysates from HeLa, A673, A549 and COS cells, using Ku80 antibody.
No image available
Immunohistochemistry Image 1: Ku80 Antibody Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Ku80 Antibody preincubated with control peptide (left) or antigen-specific peptide (right).
Immunohistochemistry Image 2: Ku80 Antibody Expand Image
Immunohistsochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localiztion, using Ku80 Antibody.
Immunofluorescence Image 1: Ku80 Antibody Expand Image
Confocal immunofluorescent analysis of HeLa cells using Ku80 Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Flow Cytometry Image 1: Ku80 Antibody Expand Image
Flow cytometric analysis of HeLa cells using Ku80 Antibody (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.