Ku70 Rabbit Polyclonal Antibody
Catalog# R1401-5
Ku70 Rabbit Polyclonal Antibody
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WB
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IHC-P
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IF-Cell
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FC
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Human
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Green monkey
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unconjugated
Overview
Product Name
Ku70 Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Synthetic peptide within N-terminal human Ku70.
Species Reactivity
Human, Green monkey
Validated Applications
WB, IHC-P, IF-Cell, FC
Molecular Weight
Predicted band size: 70 kDa
Positive Control
A549 cell lysate, HeLa cell lysate, COS-1 cell lysate, human breast cancer tissue, human tonsils tissue, HeLa.
Conjugation
unconjugated
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*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:1,000
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IF-Cell
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1:2,000
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FC
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1:1,000
Target
Function
Together, Ku70 and Ku80 make up the Ku heterodimer, which binds to DNA double-strand break ends and is required for the non-homologous end joining (NHEJ) pathway of DNA repair. It is also required for V(D)J recombination, which utilizes the NHEJ pathway to promote antigen diversity in the mammalian immune system.In addition to its role in NHEJ, Ku is also required for telomere length maintenance and subtelomeric gene silencing. Ku was originally identified when patients with systemic lupus erythematosus were found to have high levels of autoantibodies to the protein.
Background References
1. "55K isoform of CDK9 associates with Ku70 and is involved in DNA repair." Liu H., Herrmann C.H., Chiang K., Sung T.L., Moon S.H., Donehower L.A., Rice A.P. Biochem. Biophys. Res. Commun. 397:245-250(2010)
2. "Ku is a 5\'-dRP/AP lyase that excises nucleotide damage near broken ends." Roberts S.A., Strande N., Burkhalter M.D., Strom C., Havener J.M., Hasty P., Ramsden D.A. Nature 464:1214-1217(2010)
3. "Deformed epidermal autoregulatory factor-1 (DEAF1) interacts with the Ku70 subunit of the DNA-dependent protein kinase complex." Jensik P.J., Huggenvik J.I., Collard M.W. PLoS ONE 7:E33404-E33404(2012)
Sequence Similarity
Belongs to the ku70 family.
Post-translational Modification
Phosphorylation by PRKDC may enhance helicase activity. Phosphorylation of Ser-51 does not affect DNA repair.; ADP-ribosylated by PARP3.
Subcellular Location
Nucleus, Chromosome.
UNIPROT
Synonyms
5'-deoxyribose-5-phosphate lyase Ku70 antibody
5'-dRP lyase Ku70 antibody
70 kDa subunit of Ku antigen antibody
ATP dependent DNA helicase 2 subunit 1 antibody
ATP dependent DNA helicase II 70 kDa subunit antibody
ATP-dependent DNA helicase 2 subunit 1 antibody
ATP-dependent DNA helicase II 70 kDa subunit antibody
CTC box binding factor 75 kDa subunit antibody
CTC box-binding factor 75 kDa subunit antibody
CTC75 antibody
ExpandImages
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Western blot analysis of Ku70 on different lysates with Rabbit anti-Ku70 antibody (R1401-5) at 1/2,000 dilution.
Lane 1: A549 cell lysate
Lane 2: HeLa cell lysate
Lane 3: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 6 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 (R1401-5) at 1/2,000 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. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Ku70 antibody (R1401-5) at 1/1,000 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 (R1401-5) at 1/1,000 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 tonsils tissue with Rabbit anti-Ku70 antibody (R1401-5) at 1/1,000 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 (R1401-5) at 1/1,000 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. -
Immunocytochemistry analysis of HeLa cells labeling Ku70 with Rabbit anti-Ku70 antibody (R1401-5) at 1/2,000 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Ku70 antibody (R1401-5) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Flow cytometric analysis of HeLa cells labeling Ku70.
Cells were fixed and permeabilized. Then stained with the primary antibody (R1401-5, 1/1,000) (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"