Purified anti-Rad50 Antibody

Pricing & Availability
Clone
W17158C (See other available formats)
Other Names
Rad50 Double Strand Break Repair Protein, DNA Repair Protein Rad50, NBSLD
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
W17158C_PURE_Rad50_Antibody_1_073119
Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-Rad50 antibody, clone W17158C, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular Weight marker.
  • W17158C_PURE_Rad50_Antibody_1_073119
    Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-Rad50 antibody, clone W17158C, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular Weight marker.
  • W17158C_PURE_Rad50_Antibody_2_073119
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with ice-cold methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with a 1:100 dilution (5.0 µg/mL) of either purified rat IgG2a, κ isotype control antibody (Cat. No. 400602, panel A) or purified anti-Rad50 antibody (panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-rat IgG (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI and the image was captured with a 60X objective.
  • W17158C_PURE_Rad50_Antibody_3_073119
    Whole cell extracts (300 μg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.5 μg of purified rat IgG2a, κ isotype control antibody (Cat. No. 400602) or purified anti-Rad50 antibody, clone W17158C. The resulting IP fractions and whole cell extract input (5%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1.0 µg/mL (1:500 dilution) of W17158C. Lane M: Molecular Weight marker.
Cat # Size Price Quantity Avail. Save
623601 25 µg £57
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623602 100 µg £144
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Description

Rad50, along with MRE11 and NBS1, is a component of the MRN complex, which directly senses damaged DNA and displays multiple enzymatic functions required for non-homologous end joining. The MRN complex plays a critical role in the repair of double strand DNA breaks, meiotic recombination, and telomere maintenance in eukaryotes. Mutations in the individual components of the MRN complex give rise to genomic instability and are linked to multiple cancers and other pathological conditions. The Rad50 subunit binds DNA ends and retains them in close proximity to facilitate recombination events, and may directly stimulate nuclease activity of the MRE11 subunit.

Product Details
Technical Data Sheet (pdf)

Product Details

Reactivity
Human
Antibody Type
Polyclonal
Host Species
Rat
Immunogen
Partial recombinant protein corresponding to amino acid residues 1-219 of human Rad50.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP - Validated

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg per ml. For immunocytochemistry, a concentration of 5.0 μg/ml is recommended. For immunoprecipitation, the suggested use of this reagent is 2.5 µg per ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

W17158C does not recognize mouse Rad50.

 

W17158C was tested for ICC using PFA-fixed HeLa cells permeabilized with either Triton X-100 or methanol; both methods facilitated adequate staining.

Antigen Details

Structure
Rad50 is a 1,312 amino acid protein with a predicted molecular weight of 154 kD.
Distribution

Nuclear distribution/ubiquitously expressed

Function
DNA Repair, meiotic recombination, DNA damage signaling
Biology Area
Cell Biology, Cell Cycle/DNA Replication, DNA Repair/Replication
Molecular Family
Enzymes and Regulators
Antigen References
  1. Foury F, Goffeau A. 1979. Proc. Natl. Acad. Sci.USA. 76:6529-33.
  2. Liu L, et al. 2007. Nat. Cell. Biol. 9:1436-41.
  3. Lee JH, Paull TT. 2007. Oncogene. 26:7741-8.
Gene ID
10111 View all products for this Gene ID
UniProt
View information about Rad50 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 07/31/2019

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

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