Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-DNA Ligase IV/LIG4 antibody [EPR16531] - BSA and Azide free (ab232658)

Overview

  • Product name

    Anti-DNA Ligase IV/LIG4 antibody [EPR16531] - BSA and Azide free
    See all DNA Ligase IV/LIG4 primary antibodies
  • Description

    Rabbit monoclonal [EPR16531] to DNA Ligase IV/LIG4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore 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, HepG2 and HAP1 cell lysate.
  • General notes

    ab232658 is the carrier-free version of ab193353 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab232658 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product was previously labelled as DNA Ligase IV

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

    This product is a recombinant rabbit monoclonal antibody.

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

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16531
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab232658 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 Use at an assay dependent concentration. Predicted molecular weight: 103 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

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

  • 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/LIG4 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab193353).

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab193353).

References

ab232658 has not yet been referenced specifically in any publications.

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