Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)

Rabbit Recombinant Monoclonal DNA Ligase III/LIG3 antibody. Suitable for Flow Cyt (Intra), IHC-P, IP, ICC/IF, WB and reacts with Human, Mouse, Rat samples.

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Images

Flow Cytometry (Intracellular) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (AB313374), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	Flow Cyt (Intra)	IHC-P	IP	ICC/IF	WB
Human	Tested	Not recommended	Tested	Tested	Tested
Mouse	Tested	Tested	Expected	Tested	Tested
Rat	Not recommended	Tested	Expected	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -
Species Mouse	Dilution info 1/100	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -

Target data

See full target information LIG3

Function

Isoform 3 functions as a heterodimer with DNA-repair protein XRCC1 in the nucleus and can correct defective DNA strand-break repair and sister chromatid exchange following treatment with ionizing radiation and alkylating agents. Isoform 1 is targeted to mitochondria, where it functions as a DNA ligase in mitochondrial base-excision DNA repair (PubMed:10207110, PubMed:24674627).

Alternative names

DNA ligase 3, DNA ligase III, Polydeoxyribonucleotide synthase [ATP] 3, LIG3

Recommended products

Rabbit Recombinant Monoclonal DNA Ligase III/LIG3 antibody. Suitable for Flow Cyt (Intra), IHC-P, IP, ICC/IF, WB and reacts with Human, Mouse, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR27265-44
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for human IHC-P, and rat ICC and FC-Intra.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

DNA Ligase III also known as LIG3 or Lig 3 is an enzyme that plays a central role in DNA repair by catalyzing the joining of DNA strands. Mechanically it facilitates the ligation step in the DNA repair process sealing nicks in the DNA backbone. This ligase has a sizing mass of approximately 103 kDa. It is expressed widely in human cells with a high concentration in tissues with active DNA repair needs such as testes and ovaries. Ligase acts in various subcellular locations including the nucleus and mitochondria with distinct isoforms functioning in these compartments.

Biological function summary

DNA Ligase III is significant in maintaining genomic stability. It often operates as part of multi-protein complexes interacting especially with XRCC1 during base excision repair (BER). It ensures the continuity of DNA strands after damage-induced breaks are processed. LIG3 also has a pivotal function in the mitochondria where it is involved in repair and replication of mitochondrial DNA therefore preserving mitochondrial genome integrity.

Pathways

DNA Ligase III is an integral part of the DNA repair pathways particularly the base excision repair (BER) and single-strand break repair (SSBR) pathways. In the BER pathway LIG3 collaborates closely with other proteins like XRCC1 to mend damaged DNA by filling in missing nucleotides and sealing nicks. It also plays a role in non-homologous end joining (NHEJ) though this is more dominantly accomplished by DNA Ligase IV. These pathways are important for correcting oxidative damage and abasic sites that can disrupt the DNA structure.

Associated diseases and disorders

DNA Ligase III has associations with certain syndromes and cancers. Mutations or malfunctions in this enzyme can lead to ataxia-telangiectasia-like disorder owing to impaired DNA repair processes. Additionally LIG3 expression is often evaluated in the context of cancerous cells where aberrant repair processes could promote tumor progression. A connection with XRCC1 another key player in the repair machinery is critical as XRCC1 mutations may magnify the impact of LIG3 deficiencies contributing to disease pathology.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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11 product images

Flow Cytometry (Intracellular) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling DNA Ligase III/LIG3 with ab313374 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling DNA Ligase III/LIG3 with ab313374 at 1/100 (5.05 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in NIH/3T3 cells.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Immunocytochemistry/ Immunofluorescence - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling DNA Ligase III/LIG3 with ab313374 at 1/100 (5.05 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in HeLa cells.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: spleen, pancreas (PMID: 7760816).
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200, 000 dilution.
Exposure time: Lanes 1-3: 37 seconds;
Lanes 4-8: 59 seconds.
All lanes: Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374) at 1/1000 dilution
Lane 1: Rat thymus tissue lysate at 20 µg
Lane 2: Rat spleen tissue lysate at 20 µg
Lane 3: Rat pancreas tissue lysate at 20 µg
Lane 4: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 5: Mouse testis tissue lysate at 20 µg
Lane 6: Mouse thymus tissue lysate at 20 µg
Lane 7: Mouse spleen tissue lysate at 20 µg
Lane 8: Mouse pancreas tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 102,95 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling DNA Ligase III/LIG3 with ab313374 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab313374 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunoprecipitation - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
DNA Ligase III/LIG3 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab313374 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313374 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2: ab313374 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3:Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab313374 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds
All lanes: Immunoprecipitation - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374) at 1/30 dilution
All lanes: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling DNA Ligase III/LIG3 with ab313374 at 1/2000 (0.253 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab313374 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 26 seconds
All lanes: Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374) at 1/1000 dilution
Lane 1: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lane 2: Rat testis tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 102 kDa
Exposure time: 26s
Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: pancreas (PMID: 7760816).
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Exposure time: 180 seconds
All lanes: Western blot - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374) at 1/1000 dilution
Lane 1: Human testis tissue lysate at 20 µg
Lane 2: Human pancreas tissue lysate at 20 µg
Lane 3: HepG2 non-nuclear fraction at 20 µg
Lane 4: HepG2 nuclear fraction at 20 µg
Lane 5: Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate at 20 µg
Secondary
Lanes 1 - 2: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Lanes 3 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 102,95 kDa
Exposure time: 180s
Flow Cytometry (Intracellular) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling DNA Ligase III/LIG3 with ab313374 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-DNA Ligase III/LIG3 antibody [EPR27265-44] (ab313374)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling DNA Ligase III/LIG3 with ab313374 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.