Recombinant Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
![Western blot - Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-254644-anti-dna-ligase-iv-antibody-epr16531-western-blot.jpg "Western blot - 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
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 & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB 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 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.
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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| Application | Species |
|---|---|
| ICC/IF |
Human
|
| IHC-P |
Human
|
| WB |
Human
|
| 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 | (3) |
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. - 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
see all
Images
-
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)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-255436-anti-dna-ligase-iv-antibody-epr16531-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-230970-anti-dna-ligase-iv-antibody-epr16531-western-blot.jpg)
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)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-230969-anti-dna-ligase-iv-antibody-epr16531-immunohistochemistry.jpg)
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)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-230968-anti-dna-ligase-iv-antibody-epr16531-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/193/ab193353/Images/ab193353-6-benefits-of-recombinant-antibodies.png)
Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353)
Protocols
Datasheets and documents
Certificate of Compliance
References (5)
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
