XRCC5 Recombinant antibody

XRCC5 Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240301G6

Cat no : 80325-6-RR

Synonyms

KU80, CTCBF, CTC85, CTC box-binding factor 85 kDa subunit, ATP-dependent DNA helicase II 80 kDa subunit

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Validation Data Gallery

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  WB analysis using 80325-6-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 80325-6-RR (XRCC5 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 80325-6-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using XRCC5 antibody (80325-6-RR, Clone: 240301G6 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

• 

  FC experiment of MCF-7 using 80325-6-RR

  1x10^6 MCF-7 cells were intracellularly stained with 0.25 ug XRCC5 Recombinant antibody (80325-6-RR, Clone:240301G6) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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  Affinity and Kinetic Characterization of 80325-6-RR

  Biolayer interferometry (BLl) kinetic assays of 80325-6-RR against Human XRCC5 were performed. The affinity constant is 4.59 nM.

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Tested Applications

• Positive WB detected in: HeLa cells, HepG cells, HEK-293 cells, A549 cells, K-562 cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 80325-6-RR is XRCC5 Fusion Protein expressed in E. coli.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: XRCC5 fusion protein Ag9454
• 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 kDa
• GenBank Accession Number: BC019027
• Gene Symbol: XRCC5
• Gene ID (NCBI): 7520
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• 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.