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
Related conjugates and formulations
Overview
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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.
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Positive control
- WB: HeLa, Jurkat and Ramos cell lysate. IHC-P Human thymus tissue. ICC/IF: SH-SY5Y and wild-type HAP1 cells.
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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
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
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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
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Alternative Versions
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Compatible Secondaries
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Isotype control
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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.
| 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.
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| ICC/IF |
Use a concentration of 5 µg/ml.
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| WB | (3) |
1/1000 - 1/2000. Predicted molecular weight: 103 kDa.
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| Notes |
|---|
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IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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ICC/IF
Use a concentration of 5 µg/ml. |
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WB
1/1000 - 1/2000. Predicted molecular weight: 103 kDa. |
Target
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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
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Database links
- Entrez Gene: 3981 Human
- Omim: 601837 Human
- SwissProt: P49917 Human
- Unigene: 166091 Human
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Alternative names
- DNA ligase IV ATP dependent antibody
- DNA joinase antibody
- DNA ligase 4 antibody
see all
Images
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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.
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![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.
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![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.
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![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.
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![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
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Datasheet download
Certificate of Compliance
References (11)
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
