Ku80 Recombinant Rabbit Monoclonal Antibody [SC06-14]
Catalog# ET1610-40
Ku80 Recombinant Rabbit Monoclonal Antibody [SC06-14]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Green monkey
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unconjugated
Overview
Product Name
Ku80 Recombinant Rabbit Monoclonal Antibody [SC06-14]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Ku80 aa 693-732 / 732.
Species Reactivity
Human, Green monkey
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 83 kDa
Positive Control
HeLa cell lysate, MCF7 cell lysate, A549 cell lysate, COS-1 cell lysate, human colon cancer tissue, Hela, A549, SW480, human kideny tissue, human colon cancer tissue, human tonsil tissue, human breast carcinoma tissue, MCF7.
Conjugation
unconjugated
Clone Number
SC06-14
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000-1:2,000
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IF-Cell
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1:50-1:200
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IF-Tissue
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1:50-1:200
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IHC-P
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1:50-1:200
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IP
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Use at an assay dependent concentration.
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FC
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1:1,000
Target
Function
The Ku protein is localized in the nucleus and is composed of subunits referred to as Ku-70 (p70) and Ku-86 (p86) which is also known by the synonym Ku-80 or (p80). Ku was first described as an autoantigen to which antibodies were produced in a patient with scleroderma polymyositis overlap syndrome, and was later found in the sera of patients with other rheumatic diseases. Both subunits of the Ku protein have been cloned, and a number of functions have been proposed for Ku, including cell signaling, DNA replication and transcriptional activation. Ku is involved in Pol II-directed transcription by virtue of its DNA binding activity, serving as the regulatory component of the DNA-associated protein kinase that phosphorylates Pol II and transcription factor Sp. Ku proteins also activate transcription from the U1 small nuclear RNA and the human transferrin receptor gene promoters. A Ku-related protein designated the enhancer 1 binding factor (E1BF), composed of two subunits, has been identified as a positive regulator of RNA polymerase I transcription initiation.
Background References
1. Ho SR et al. RNF144A, an E3 ubiquitin ligase for DNA-PKcs, promotes apoptosis during DNA damage. Proc Natl Acad Sci U S A 111:E2646-55 (2014).
2. Grundy GJ et al. APLF promotes the assembly and activity of non-homologous end joining protein complexes. EMBO J 32:112-25 (2013).
Sequence Similarity
Belongs to the ku80 family.
Post-translational Modification
ADP-ribosylated by PARP3.; Phosphorylated on serine residues. Phosphorylation by PRKDC may enhance helicase activity.; Sumoylated.; Ubiquitinated by RNF8 via 'Lys-48'-linked ubiquitination following DNA damage, leading to its degradation and removal from DNA damage sites. Ubiquitinated by RNF138, leading to remove the Ku complex from DNA breaks.
Subcellular Location
Nucleus, Chromosome.
UNIPROT
Synonyms
86 kDa subunit of Ku antigen antibody
ATP dependent DNA helicase 2 subunit 2 antibody
ATP dependent DNA helicase II 80 kDa subunit antibody
ATP dependent DNA helicase II 86 Kd subunit antibody
ATP dependent DNA helicase II antibody
ATP-dependent DNA helicase 2 subunit 2 antibody
ATP-dependent DNA helicase II 80 kDa subunit antibody
CTC box binding factor 85 kDa antibody
CTC box-binding factor 85 kDa subunit antibody
CTC85 antibody
ExpandImages
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Western blot analysis of Ku80 on different lysates with Rabbit anti-Ku80 antibody (ET1610-40) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: A549 cell lysate
Lane 4: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 83 kDa
Observed band size: 83 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-40) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of Ku80 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Ku80 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Ku80 in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Ku80 antibody (ET1610-40) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-40) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Ku80 antibody (ET1610-40) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-40) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human kideny tissue with Rabbit anti-Ku80 antibody (ET1610-40) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-40) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Ku80 antibody (ET1610-40) at 1/50 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-40) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of MCF7 cells labeling Ku80.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-40, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"