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AB198587

Alexa Fluor® 647 Anti-Ku80 antibody [EPR3468]

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(1 Publication)

Rabbit Recombinant Monoclonal Ku80 antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 1 publication.

View Alternative Names

G22P2, XRCC5, X-ray repair cross-complementing protein 5, 86 kDa subunit of Ku antigen, ATP-dependent DNA helicase 2 subunit 2, ATP-dependent DNA helicase II 80 kDa subunit, CTC box-binding factor 85 kDa subunit, DNA repair protein XRCC5, Ku80, Ku86, Lupus Ku autoantigen protein p86, Nuclear factor IV, Thyroid-lupus autoantigen, X-ray repair complementing defective repair in Chinese hamster cells 5 (double-strand-break rejoining), CTC85, CTCBF, TLAA

2 Images
Flow Cytometry - Alexa Fluor® 647 Anti-Ku80 antibody [EPR3468] (AB198587)
  • Flow Cyt

Lab

Flow Cytometry - Alexa Fluor® 647 Anti-Ku80 antibody [EPR3468] (AB198587)

Overlay histogram showing HeLa cells stained with ab198587 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab198587, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Ku80 antibody [EPR3468] (AB198587)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Ku80 antibody [EPR3468] (AB198587)

ab198587 staining Ku80 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 ab198587 at 1/100 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 HeLacells fixed with 4% formaldehyde (10 min).

  • HRP

    HRP Anti-Ku80 antibody [EPR3468]

  • 578 PE

    PE Anti-Ku80 antibody [EPR3468]

  • 519 Alexa Fluor® 488

    Alexa Fluor® 488 Anti-Ku80 antibody [EPR3468]

  • Unconjugated

    Anti-Ku80 antibody [EPR3468]

  • Carrier free

    Anti-Ku80 antibody [EPR3468] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3468

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human

Applications

ICC/IF, Flow Cyt (Intra)

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"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "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>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/50", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/alexa-fluor-647-rabbit-igg-monoclonal-epr25a-isotype-control-ab199093'>ab199093</a> - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.</p>" }, "Mouse": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "1/100", "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>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "1/50", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/alexa-fluor-647-rabbit-igg-monoclonal-epr25a-isotype-control-ab199093'>ab199093</a> - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.</p>" }, "Rat": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "1/100", "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>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "1/50", "FlowCytIntra-species-notes": "<p><a href='/en-us/products/primary-antibodies/alexa-fluor-647-rabbit-igg-monoclonal-epr25a-isotype-control-ab199093'>ab199093</a> - Rabbit monoclonal IgG (Alexa Fluor®; 647), is suitable for use as an isotype control with this antibody.</p>" } } }
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Product details

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.

The Ku80 protein also known as Ku p70 Ku antigen p70 and by gene identifiers such as Ku80 hum39 and 5c5 functions critically in DNA repair processes. With a molecular mass of approximately 80 kDa Ku80 is ubiquitously expressed in mammalian cells. It is an important component of the Ku heterodimer working with its partner Ku70 to detect and bind DNA double-strand breaks (DSBs). This action initiates repair by aligning with DNA ends to prevent break processing and sustain genome integrity.
Biological function summary

The Ku80 protein executes an important role in maintaining cellular stability by participating in the non-homologous end joining (NHEJ) pathway. Ku80 forms a complex with DNA-PKcs (DNA-dependent protein kinase catalytic subunit) which activates the kinase to phosphorylate itself and other proteins facilitating the repair of DSBs. Its expression is significant in tissues that undergo high rates of cellular turnover and are experienced with DNA damage where the DNA repair mechanisms are important for regular cell cycle progression.

Pathways

DNA repair and V(D)J recombination are principal domains where Ku80 shows its influence. In the context of DNA repair Ku80's partnership with DNA-PKcs as part of the NHEJ repair pathway is fundamental in fixing DNA damage from environmental insults or normal cellular metabolism. In the immune development process through V(D)J recombination Ku80 partners with other proteins like the Artemis complex to perform programmed DNA rearrangements required for the diversity of immune receptors.

Ku80 has a significant relation to cancer and immune deficiencies. Defects in Ku80 function can lead to increased sensitivity to ionizing radiation and a higher propensity for cancer development because of impaired DNA repair capabilities. Moreover in immune-related disorders inadequate Ku80 performance affects the maturation of lymphocytes leading to immune system malfunction. Its malfunctioning connection with proteins like DNA-PKcs and Artemis highlights its critical roles in both tumor suppression and immune system competence.
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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

Single-stranded DNA-dependent ATP-dependent helicase that plays a key role in DNA non-homologous end joining (NHEJ) by recruiting DNA-PK to DNA (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). Required for double-strand break repair and V(D)J recombination (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). Also has a role in chromosome translocation (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). The DNA helicase II complex binds preferentially to fork-like ends of double-stranded DNA in a cell cycle-dependent manner (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). It works in the 3'-5' direction (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). During NHEJ, the XRCC5-XRRC6 dimer performs the recognition step : it recognizes and binds to the broken ends of the DNA and protects them from further resection (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). Binding to DNA may be mediated by XRCC6 (PubMed : 11493912, PubMed : 12145306, PubMed : 7957065, PubMed : 8621488). The XRCC5-XRRC6 dimer acts as a regulatory subunit of the DNA-dependent protein kinase complex DNA-PK by increasing the affinity of the catalytic subunit PRKDC to DNA by 100-fold (PubMed : 11493912, PubMed : 12145306, PubMed : 20383123, PubMed : 7957065, PubMed : 8621488). The XRCC5-XRRC6 dimer is probably involved in stabilizing broken DNA ends and bringing them together (PubMed : 12145306, PubMed : 20383123, PubMed : 7957065, PubMed : 8621488). The assembly of the DNA-PK complex to DNA ends is required for the NHEJ ligation step (PubMed : 12145306, PubMed : 20383123, PubMed : 7957065, PubMed : 8621488). The XRCC5-XRRC6 dimer probably also acts as a 5'-deoxyribose-5-phosphate lyase (5'-dRP lyase), by catalyzing the beta-elimination of the 5' deoxyribose-5-phosphate at an abasic site near double-strand breaks (PubMed : 20383123). XRCC5 probably acts as the catalytic subunit of 5'-dRP activity, and allows to 'clean' the termini of abasic sites, a class of nucleotide damage commonly associated with strand breaks, before such broken ends can be joined (PubMed : 20383123). The XRCC5-XRRC6 dimer together with APEX1 acts as a negative regulator of transcription (PubMed : 8621488). In association with NAA15, the XRCC5-XRRC6 dimer binds to the osteocalcin promoter and activates osteocalcin expression (PubMed : 12145306). As part of the DNA-PK complex, involved in the early steps of ribosome assembly by promoting the processing of precursor rRNA into mature 18S rRNA in the small-subunit processome (PubMed : 32103174). Binding to U3 small nucleolar RNA, recruits PRKDC and XRCC5/Ku86 to the small-subunit processome (PubMed : 32103174). Plays a role in the regulation of DNA virus-mediated innate immune response by assembling into the HDP-RNP complex, a complex that serves as a platform for IRF3 phosphorylation and subsequent innate immune response activation through the cGAS-STING pathway (PubMed : 28712728).
See full target information XRCC5
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

eLife 7: PubMed29985131

2018

PARP1-dependent recruitment of the FBXL10-RNF68-RNF2 ubiquitin ligase to sites of DNA damage controls H2A.Z loading.

Applications

Unspecified application

Species

Unspecified reactive species

Gergely Rona,Domenico Roberti,Yandong Yin,Julia K Pagan,Harrison Homer,Elizabeth Sassani,Andras Zeke,Luca Busino,Eli Rothenberg,Michele Pagano
View all publications
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