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  1. Link

    dna-ligase-ivlig4-antibody-epr16531-ab193353.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA Synthesis DNA Ligases
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (4) Submit a question References (11)

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

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

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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. This information is proprietary to Abcam and/or its suppliers.

  • 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 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.

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

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Synthesis
    • DNA Ligases

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab225387)
    • Anti-DNA Ligase IV/LIG4 antibody [EPR16531] - BSA and Azide free (ab232658)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Ramos whole cell lysate (ab3955)
    • Jurkat whole cell lysate (ab7899)

Applications

The Abpromise guarantee

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 (4)
1/1000 - 1/2000. Predicted molecular weight: 103 kDa.
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.
  • Target information above from: UniProt accession P49917 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3981 Human
    • Omim: 601837 Human
    • SwissProt: P49917 Human
    • Unigene: 166091 Human
    • 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.

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

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (11)

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

    ab193353 has been referenced in 11 publications.

    • Burgess JT  et al. Barrier-to-autointegration-factor (Banf1) modulates DNA double-strand break repair pathway choice via regulation of DNA-dependent kinase (DNA-PK) activity. Nucleic Acids Res 49:3294-3307 (2021). PubMed: 33660778
    • Silva B  et al. TERRA transcription destabilizes telomere integrity to initiate break-induced replication in human ALT cells. Nat Commun 12:3760 (2021). PubMed: 34145295
    • Liu X  et al. Chromosomal aberration arises during somatic reprogramming to pluripotent stem cells. Cell Div 15:12 (2020). PubMed: 33292330
    • Rother MB  et al. CHD7 and 53BP1 regulate distinct pathways for the re-ligation of DNA double-strand breaks. Nat Commun 11:5775 (2020). PubMed: 33188175
    • Fan Z  et al. Checkpoint kinase-1 inhibition and etoposide exhibit a strong synergistic anticancer effect on chronic myeloid leukemia cell line K562 by impairing homologous recombination DNA damage repair. Oncol Rep 44:2152-2164 (2020). PubMed: 32901871
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U20S)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    50 µg
    Specification
    U20S
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 19 2022

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

    Excellent
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 06 2020

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

    Poor
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    靖友 湯徳

    Verified customer

    Submitted Sep 21 2016

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

    Poor
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 26 2016

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