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

     

    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

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 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

  • 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.

  • 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.

  • 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

  • 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.

  • 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

This product has been referenced in:

See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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

靖友 湯徳

Verified customer

Submitted Sep 21 2016

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

Abcam user community

Verified customer

Submitted Jul 26 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up