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AB225387

Alexa Fluor® 647 Anti-DNA Ligase IV/LIG4 antibody [EPR16531]

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Rabbit Recombinant Monoclonal DNA Ligase IV/LIG4 antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF and reacts with Human samples.

View Alternative Names

DNA ligase 4, DNA ligase IV, Polydeoxyribonucleotide synthase [ATP] 4, LIG4

1 Images
Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (AB225387)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (AB225387)

ab225387 staining DNA Ligase IV/LIG4 in HeLa cells. The cells were fixed with 100% methanol (5 min), 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 overnight at +4°C with ab225387 at 1/1000 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

  • Unconjugated

    Anti-DNA Ligase IV/LIG4 antibody [EPR16531]

  • Carrier free

    Anti-DNA Ligase IV/LIG4 antibody [EPR16531] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR16531

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human

Applications

ICC/IF

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/1000", "ICCIF-species-notes": "<p>This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)</p>" } } }
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Product details

This conjugated antibody has been KO validated based on the results obtained with the unconjugated clone: Anti-DNA Ligase IV/LIG4 antibody [EPR16531] (ab193353).

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
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|Store in the dark
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Supplementary information

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

DNA Ligase IV also known as LIG4 or DNA Ligase 4 is an enzyme with a molecular mass of approximately 103 kDa. It is known to play a vital role in the DNA repair process by catalyzing the final step of DNA double-strand breaks where it joins two DNA strands in a phosphodiester bond. DNA Ligase IV is widely expressed in human tissues and its activity is key in the non-homologous end joining (NHEJ) pathway which repairs double-strand breaks in DNA. This protein forms a multi-protein complex with its cofactor XRCC4 and sometimes with other accessory proteins for efficient functioning.
Biological function summary

The significance of DNA Ligase IV in maintaining genomic stability makes it essential for proper cellular functions. DNA Ligase IV is a part of the NHEJ repair complex and works in close association with proteins such as XRCC4 and XLF-Cernunnos which stabilize the complex. It orchestrates the repair of DNA damage that could otherwise lead to mutations or chromosomal translocations. Furthermore LIG4 is involved in V(D)J recombination a process critical for immune system development by creating diversity in immunoglobulins and T cell receptors.

Pathways

DNA Ligase IV plays a significant role in the NHEJ pathway which is one of the two main pathways for repairing DNA double-strand breaks. This pathway provides a rapid and essential means of maintaining DNA integrity. DNA-PKcs a kinase activates the NHEJ pathway and works closely with LIG4. Additionally LIG4 takes part in V(D)J recombination orchestrating the proper joining of DNA segments important for developing a functional immune system. The protein Ku is also essential in these pathways acting as a scaffold for the entire repair machinery.

Defects or mutations in DNA Ligase IV can lead to severe health issues such as LIG4 syndrome a condition characterized by immunodeficiency and developmental delay and increased radiation sensitivity. Another disorder associated is leukemia as faulty NHEJ can result in chromosomal translocations leading to cancer. The interaction with XRCC4 is necessary for its DNA repair tasks and mutations affecting this interaction can exacerbate the disorder's severity. Understanding the role of DNA Ligase IV in these contexts provides insight into potential therapeutic targets for managing such conditions.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

DNA ligase involved in DNA non-homologous end joining (NHEJ); required for double-strand break (DSB) repair and V(D)J recombination (PubMed : 12517771, PubMed : 17290226, PubMed : 23523427, PubMed : 29980672, PubMed : 33586762, PubMed : 8798671, PubMed : 9242410, PubMed : 9809069). Catalyzes the NHEJ ligation step of the broken DNA during DSB repair by resealing the DNA breaks after the gap filling is completed (PubMed : 12517771, PubMed : 17290226, PubMed : 9242410, PubMed : 9809069). Joins single-strand breaks in a double-stranded polydeoxynucleotide in an ATP-dependent reaction (PubMed : 12517771, PubMed : 17290226, PubMed : 9242410, PubMed : 9809069). LIG4 is mechanistically flexible : it can ligate nicks as well as compatible DNA overhangs alone, while in the presence of XRCC4, it can ligate ends with 2-nucleotides (nt) microhomology and 1-nt gaps (PubMed : 17290226). Forms a subcomplex with XRCC4; the LIG4-XRCC4 subcomplex is responsible for the NHEJ ligation step and XRCC4 enhances the joining activity of LIG4 (PubMed : 9242410, PubMed : 9809069). 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 (PubMed : 10854421). LIG4 regulates nuclear localization of XRCC4 (PubMed : 24984242).
See full target information LIG4
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Product promise

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