Overview

  • Product nameAnti-Ku70 antibody
    See all Ku70 primary antibodies
  • Description
    Rabbit polyclonal to Ku70
  • Tested applicationsIP, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 550 to the C-terminus of Human Ku70.

    (Peptide available as ab92421.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Irradiated HeLa; HepG2; Jurkat; MCF7; U2OS.

Properties

Applications

Our Abpromise guarantee covers the use of ab83501 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 70 kDa).Can be blocked with Human Ku70 peptide (ab92421).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionSingle stranded DNA-dependent ATP-dependent helicase. Has a role in chromosome translocation. The DNA helicase II complex binds preferentially to fork-like ends of double-stranded DNA in a cell cycle-dependent manner. It works in the 3'-5' direction. Binding to DNA may be mediated by XRCC6. Involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination. The XRCC5/6 dimer acts as regulatory subunit of the DNA-dependent protein kinase complex DNA-PK by increasing the affinity of the catalytic subunit PRKDC to DNA by 100-fold. The XRCC5/6 dimer is probably involved in stabilizing broken DNA ends and bringing them together. The assembly of the DNA-PK complex to DNA ends is required for the NHEJ ligation step. Required for osteocalcin gene expression. Probably also acts as a 5'-deoxyribose-5-phosphate lyase (5'-dRP lyase), by catalyzing the beta-elimination of the 5' deoxyribose-5-phosphate at an abasic site near double-strand breaks. 5'-dRP lyase activity allows to 'clean' the termini of abasic sites, a class of nucleotide damage commonly associated with strand breaks, before such broken ends can be joined. The XRCC5/6 dimer together with APEX1 acts as a negative regulator of transcription.
  • Sequence similaritiesBelongs to the ku70 family.
    Contains 1 Ku domain.
    Contains 1 SAP domain.
  • Developmental stageExpression does not increase during promyelocyte differentiation.
  • Post-translational
    modifications
    Phosphorylation by PRKDC may enhance helicase activity. Phosphorylation of Ser-51 does not affect DNA repair.
  • Cellular localizationNucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • 5''-deoxyribose-5-phosphate lyase Ku70 antibody
    • 5''-dRP lyase Ku70 antibody
    • 70 kDa subunit of Ku antigen antibody
    • ATP dependent DNA helicase 2 subunit 1 antibody
    • ATP dependent DNA helicase II 70 kDa subunit antibody
    • ATP-dependent DNA helicase 2 subunit 1 antibody
    • ATP-dependent DNA helicase II 70 kDa subunit antibody
    • CTC box binding factor 75 kDa subunit antibody
    • CTC box-binding factor 75 kDa subunit antibody
    • CTC75 antibody
    • CTCBF antibody
    • DNA repair protein XRCC6 antibody
    • G22P1 antibody
    • Ku 70 antibody
    • Ku autoantigen p70 subunit antibody
    • Ku autoantigen, 70kDa antibody
    • Ku p70 antibody
    • Ku70 antibody
    • Ku70 DNA binding component of DNA-dependent proteinkinase complex (thyroid autoantigen 70 kDa antibody
    • Kup70 antibody
    • Lupus Ku autoantigen protein p70 antibody
    • ML8 antibody
    • Thyroid autoantigen 70kD (Ku antigen) antibody
    • Thyroid autoantigen antibody
    • Thyroid lupus autoantigen antibody
    • Thyroid lupus autoantigen p70 antibody
    • Thyroid-lupus autoantigen antibody
    • TLAA antibody
    • X ray repair complementing defective repair in Chinese hamster cells 6 antibody
    • X-ray repair complementing defective repair in Chinese hamster cells 6 antibody
    • X-ray repair cross-complementing protein 6 antibody
    • XRCC 6 antibody
    • XRCC6 antibody
    • XRCC6_HUMAN antibody
    see all

Anti-Ku70 antibody images

  • Ku70 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Ku70 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab83501.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 70kDa; Ku70

  • All lanes : Anti-Ku70 antibody (ab83501) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Irradiated Hela (Control for ATM phospho S1981) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 73 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute
  • ICC/IF image of ab83501 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab83501, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2, MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
  • IHC image of Ku70 staining in human kidney carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab83501, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

References for Anti-Ku70 antibody (ab83501)

This product has been referenced in:
  • Changou CA  et al. Arginine starvation-associated atypical cellular death involves mitochondrial dysfunction, nuclear DNA leakage, and chromatin autophagy. Proc Natl Acad Sci U S A 111:14147-52 (2014). ICC/IF . Read more (PubMed: 25122679) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa cells - cervical epithelial cancer)
Specification HeLa cells - cervical epithelial cancer
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% Triton/TBS
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Jan 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells - cervical epithelial cancer)
Loading amount 10 µg
Specification HeLa cells - cervical epithelial cancer
Gel Running Conditions Reduced Denaturing (4-20% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Dec 09 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"