Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)

Reviews (3) Submit a question References (5)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16531] to DNA Ligase IV/LIG4
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-DNA Ligase IV/LIG4 antibody [EPR16531]
    See all DNA Ligase IV/LIG4 primary antibodies

  • Description

    Rabbit monoclonal [EPR16531] to DNA Ligase IV/LIG4

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human DNA Ligase IV/LIG4 aa 500-650. The exact sequence is proprietary.
    Database link: P49917

  • Positive control

    • WB: HeLa, Jurkat and Ramos cell lysate. IHC-P Human thymus tissue. ICC/IF: SH-SY5Y and wild-type HAP1 cells.

  • General notes

     This product was previously labelled as DNA Ligase IV

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR16531

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab193353 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
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB 1/1000 - 1/2000. Predicted molecular weight: 103 kDa.

Target

  • Function

    Efficiently joins single-strand breaks in a double-stranded polydeoxynucleotide in an ATP-dependent reaction. Involved in DNA non-homologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination. The LIG4-XRCC4 complex is responsible for the NHEJ ligation step, and XRCC4 enhances the joining activity of LIG4. Binding of the LIG4-XRCC4 complex to DNA ends is dependent on the assembly of the DNA-dependent protein kinase complex DNA-PK to these DNA ends.

  • Tissue specificity

    Testis, thymus, prostate and heart.

  • Involvement in disease

    LIG4 syndrome
    Severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-positive with sensitivity to ionizing radiation

  • Sequence similarities

    Belongs to the ATP-dependent DNA ligase family.
    Contains 2 BRCT domains.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • DNA ligase IV ATP dependent antibody
      • DNA joinase antibody
      • DNA ligase 4 antibody
      • DNA ligase IV antibody
      • DNA repair enzyme antibody
      • DNLI4_HUMAN antibody
      • LIG 4 antibody
      • LIG4 antibody
      • LIG4S antibody
      • Ligase IV antibody
      • Ligase IV DNA ATP dependent antibody
      • Polydeoxyribonucleotide synthase [ATP] 4 antibody
      • Polydeoxyribonucleotide synthase 4 antibody
      • Polydeoxyribonucleotide synthase antibody
      • Polynucleotide ligase antibody
      • Sealase antibody
      see all

    Images

    • Western blot - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      Western blot - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: DNA Ligase IV/LIG4 knockout HAP1 cell lysate (20 µg)
      Lane 3: HepG2 cell lysate (20 µg)
      Lane 4: HeLa cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab193353 observed at 103 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab193353 was shown to specifically react with DNA Ligase IV/LIG4 when DNA ligase IV knockout samples were used. Wild-type and DNA Ligase IV/LIG4 knockout samples were subjected to SDS-PAGE. ab193353 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)

      ab193353 staining DNA Ligase IV/LIG4 in wild-type HAP1 cells (top panel) and LIG4 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab193353 at 5μg/ml dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Western blot - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      Western blot - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      All lanes : Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 103 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)

      Immunohistochemical analysis of paraffin embedded Human thymus tissue sections labeling DNA Ligase IV/LIG4 using ab193353 at a 1/50 dilution. A ready to use HRP Polymer for Rabbit IgG was used as the secondary. Hematoxylin counterstain.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
      Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)

      Immunofluorescent analysis of 4% paraformaldehyde fixed SH-SY5Y cells labeling DNA Ligase IV/LIG4 using ab193353 at a 1/50 dilution. A Goat anti rabbit IgG (Alexa Fluor®488) (ab150077) was used as the secondary at a 1/400 dilution. Counterstain DAPI. Cells were permeabilized using 0.1% Triton X-100.

    References (5)

    Publishing research using ab193353? Please let us know so that we can cite the reference in this datasheet.

    ab193353 has been referenced in 5 publications.

    • Vítor AC  et al. Single-molecule imaging of transcription at damaged chromatin. Sci Adv 5:eaau1249 (2019). PubMed: 30662944
    • Xing M & Oksenych V Genetic interaction between DNA repair factors PAXX, XLF, XRCC4 and DNA-PKcs in human cells. FEBS Open Bio 9:1315-1326 (2019). PubMed: 31141305
    • Han J  et al. BRCA2 antagonizes classical and alternative nonhomologous end-joining to prevent gross genomic instability. Nat Commun 8:1470 (2017). PubMed: 29133916
    • Arnoult N  et al. Regulation of DNA repair pathway choice in S and G2 phases by the NHEJ inhibitor CYREN. Nature 549:548-552 (2017). PubMed: 28959974
    • McManus KJ & Hendzel MJ ATM-dependent DNA damage-independent mitotic phosphorylation of H2AX in normally growing mammalian cells. Mol Biol Cell 16:5013-25 (2005). PubMed: 16030261

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-DNA Ligase IV/LIG4 antibody [EPR16531]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human coronary artery endothelial cells HCAEC)
    Gel Running Conditions
    Reduced Denaturing (4-20% gradient)
    Loading amount
    15 µg
    Specification
    Human coronary artery endothelial cells HCAEC
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 06 2020

    Western blot abreview for Anti-DNA Ligase IV/LIG4 antibody [EPR16531]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (MDCK)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    75 µg
    Specification
    MDCK
    Blocking step
    ECL Prime Blocking Reagent as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 23°C
    Read More

    靖友 湯徳

    Verified customer

    Submitted Sep 21 2016

    Western blot abreview for Anti-DNA Ligase IV/LIG4 antibody [EPR16531]

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Abelsons)
    Gel Running Conditions
    Reduced Denaturing (8% SDS-Page)
    Loading amount
    1e+006 cells
    Specification
    Abelsons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 26 2016

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