Overview

  • Product name

    Anti-Ku70 antibody [EPR4027]
    See all Ku70 primary antibodies
  • Description

    Rabbit monoclonal [EPR4027] to Ku70
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Ku70 aa 500-600. The exact sequence is proprietary.
    Database link: P12956

  • Positive control

    • WB: A549, 293T, A431 , and HeLa lysates. IHC-P: human colon carcinoma, tonsil and testis tissue. ICC/IF: HeLa cells. FC: HeLa cells. IP: HeLa cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92450 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
WB 1/1000 - 1/10000. Predicted molecular weight: 70 kDa.
IP 1/20.
IHC-P 1/100 - 1/250.

Antigen retrieval andthe use of an HRP/AP polymerized secondary antibody is recommended for enhanced staining.

See IHC antigen retrieval protocols.

 

Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.

Target

  • Function

    Single 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 similarities

    Belongs to the ku70 family.
    Contains 1 Ku domain.
    Contains 1 SAP domain.
  • Developmental stage

    Expression 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 localization

    Nucleus. 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

Images

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ku70 with purified ab92450 at 1/250 dilution (0.46 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon carcinoma tissue sections labeling Ku70 with purified ab92450 at 1/250 dilution (0.46 µg/ml). Heat mediated antigen retrieval using Bond� Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • All lanes : Anti-Ku70 antibody [EPR4027] (ab92450) at 1/1000 dilution ((unpurified))

    Lane 1 : HeLa cell lysate
    Lane 2 : 293T cell lysate
    Lane 3 : A549 cell lysate
    Lane 4 : A431 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 70 kDa

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Ku70 with purified ab92450 at 1/20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • ab92450 (purified ) at 1/20 dilution (0.5ug) immunoprecipitating Ku70 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab92450 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92450 in HeLa whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP)�(ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Immunofluorescence staining of HeLa cells using unpurified ab92450 at 1/100 dilution.

  • Overlay histogram showing HeLa cells stained with unpurified ab92450 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92450, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Paraffin embedded Human testis tissue (A) or Human tonsil tissue (B) were labelled with unpurified ab92450 at 1/100 dilution.

References

This product has been referenced in:

  • Lin Y  et al. Effects of combined radiosurgery and temozolomide therapy on epidermal growth factor receptor and variant III in glioblastoma multiforme. Oncol Lett 15:5751-5759 (2018). Read more (PubMed: 29563997) »
  • Lobbardi R  et al. TOX Regulates Growth, DNA Repair, and Genomic Instability in T-cell Acute Lymphoblastic Leukemia. Cancer Discov 7:1336-1353 (2017). Read more (PubMed: 28974511) »
See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
U2OS
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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Verified customer

Submitted Nov 04 2015

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