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
Product nameAnti-DNA Ligase IV/LIG4 antibody [EPR16531]
See all DNA Ligase IV/LIG4 primary antibodies
DescriptionRabbit monoclonal [EPR16531] to DNA Ligase IV/LIG4
Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Human
Recombinant fragment within Human DNA Ligase IV/LIG4 aa 500-650. The exact sequence is proprietary.
Database link: P49917
- WB: HeLa, Jurkat and Ramos cell lysate. IHC-P Human thymus tissue. ICC/IF: SH-SY5Y and wild-type HAP1 cells.
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
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.
Storage instructionsShipped 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 bufferpH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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.|
FunctionEfficiently 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 specificityTestis, thymus, prostate and heart.
Involvement in diseaseLIG4 syndrome
Severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-negative/NK-cell-positive with sensitivity to ionizing radiation
Sequence similaritiesBelongs to the ATP-dependent DNA ligase family.
Contains 2 BRCT domains.
- Information by UniProt
- DNA ligase IV ATP dependent antibody
- DNA joinase antibody
- DNA ligase 4 antibody
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.
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.
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