XRCC5 Monoclonal antibody

XRCC5 Monoclonal Antibody for IF, IHC, IP, WB, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human, mouse, rat and More (1)

Applications

WB, IP, IHC, IF, ELISA

Conjugate

Unconjugated

Publications

3

CloneNo.

2G5E7

Cat no : 66546-1-Ig

Synonyms

86 kDa subunit of Ku antigen, CTC85, CTCBF, DNA repair protein XRCC5, G22P2, KARP 1, KARP1, KU80, Ku86, KUB2, NFIV, Nuclear factor IV, Thyroid lupus autoantigen, TLAA, XRCC5

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Validation Data Gallery

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  WB analysis using 66546-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66546-1-Ig (XRCC5 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HSC-T6 using 66546-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66546-1-Ig (XRCC5 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 66546-1-Ig

  IP result of anti-XRCC5 (IP:66546-1-Ig, 5ug; Detection:66546-1-Ig 1:20000) with HeLa cells lysate 3200 ug.

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  IP experiment of HeLa using 66546-1-Ig

  IP result of anti-XRCC5(IP:66546-1-Ig, 5ug; Detection:66546-1-Ig 1:40000) with HeLa cells lysate 640 ug.

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  IHC staining of human lung cancer using 66546-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66546-1-Ig (XRCC5 antibody) at dilution of 1:1000 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66546-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66546-1-Ig (XRCC5 antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66546-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66546-1-Ig (XRCC5 antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 66546-1-Ig

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66546-1-Ig (XRCC5 antibody) at dilution of 1:1000 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 66546-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using 66546-1-Ig (XRCC5 antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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Tested Applications

• Positive WB detected in: HeLa cells, HSCT6 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lung cancer tissue, human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:5000-1:50000 for WB
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• WB: See 1 publications below
• IF: See 2 publications below

Product Information

66546-1-Ig targets XRCC5 in WB, IP, IHC, IF, ELISA applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: XRCC5 fusion protein Ag9512
• Full Name: X-ray repair complementing defective repair in Chinese hamster cells 5 (double-strand-break rejoining)
• Calculated molecular weight: 732 aa, 83 kDa
• Observed molecular weight: 80-83 kDa
• GenBank accession number: BC019027
• Gene symbol: XRCC5
• Gene ID (NCBI): 7520
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

There are at least two pathways for eukaryotes to repair DNA double-strand breaks: homologous recombination and nonhomologous end joining(NHEJ). The core NHEJ machinery includes XRCC4, DNA ligase IV and the DNA-dependent protein kinase complex, which consists of the DNA end-binding XRCC5/XRCC6 heterodimer and the catalytic subunit PRKDC. The heterdimer of XRCC5/XRCC6 enhanced teh affinity of the catalytic subunit PRKDC to DNA by 100-fold. Once the XRCC5/6 dimer association with NAA15, it can bind to the osteocalcin promoter and activate osteocalcin expression. The XRCC5/6 dimer acts as a negative regulator of transcription when together with APEX1. Some publised papers indicated that the MW of XRCC5 is 86kDa, while more papers suggested that XRCC5 is a 80kDa protein, as it was firstly introducted in publication. Thus, Ku80 and Ku86 are the same protein.

Publications

Species	Application	Title
human	WB	Cancer Lett; ITGA2 overexpression inhibits DNA repair and confers sensitivity to radiotherapies in pancreatic cancer; Authors - Chen Zhou
human	IF	iScience; WASH interacts with Ku to regulate DNA double-stranded break repair.; Authors - Tao Wang
human	IF	Cell Biol Int; Comparison of DNA stability and its related genes of neurons derived from induced pluripotent stem cells and primary retinal neurons.; Authors - Na Yu