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Ku70 Recombinant Rabbit Monoclonal Antibody [JM61-31]

Usd: 385

Specification

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Catalog# ET1703-95

Ku70 Recombinant Rabbit Monoclonal Antibody [JM61-31]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

Ku70 Recombinant Rabbit Monoclonal Antibody [JM61-31]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Ku70 aa 521-564 / 609.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 70 kDa

Positive Control

HeLa cell lysate, HeLa, human tonsil tissue.

Conjugation

unconjugated

Clone Number

JM61-31

RRID

AB_3070449

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

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:500

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:1,000

  • FC

  • 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. Mohni KN et al. ATR pathway inhibition is synthetically lethal in cancer cells with ERCC1 deficiency. Cancer Res 74:2835-45 (2014).

2. Parvathaneni S et al. Human RECQ1 interacts with Ku70/80 and modulates DNA end-joining of double-strand breaks. PLoS One 8:e62481 (2013).

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

SwissProt: P12956 Human

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

Expand

Images

  • Immunocytochemistry analysis of HeLa cells labeling Ku70 with Rabbit anti-Ku70 antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) at 1/500 dilution.<br /><br />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 (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Ku70 with Rabbit anti-Ku70 antibody (ET1703-95) at 1/500 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 (ET1703-95) at 1/500 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Ku70 antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) 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 tonsil tissue with Rabbit anti-Ku70 antibody (ET1703-95) 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 (ET1703-95) 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 tonsil tissue with Rabbit anti-Ku70 antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) 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 tonsil tissue with Rabbit anti-Ku70 antibody (ET1703-95) 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 (ET1703-95) 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.

  • Flow cytometric analysis of HeLa cells labeling Ku70.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Ku70.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-95, 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).

  • Western blot analysis of Ku70 on HeLa cell lysate with Rabbit anti-Ku70 antibody (<a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>) at 1/2,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 10 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1703-95" style="font-weight: bold;text-decoration: underline;">ET1703-95</a>, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 70 kDa<br />Observed band size: 70 kDa

    Western blot analysis of Ku70 on HeLa cell lysate with Rabbit anti-Ku70 antibody (ET1703-95) at 1/2,000 dilution.

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 10 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1703-95, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 70 kDa
    Observed band size: 70 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"