Recombinant
RabMAb

Anti-Rad50 antibody [EPR3466(2)] (ab124682)

Overview

  • Product name
    Anti-Rad50 antibody [EPR3466(2)]
    See all Rad50 primary antibodies
  • Description
    Rabbit monoclonal [EPR3466(2)] to Rad50
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide, corresponding to residues in Human Rad50.

  • Positive control
    • K562, MOLT4, Jurkat and MCF7 cell lysates.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124682 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/80.
ICC/IF 1/100.
WB 1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 154 kDa).

Target

  • Function
    Component of the MRN complex, which plays a central role in double-strand break (DSB) repair, DNA recombination, maintenance of telomere integrity and meiosis. The complex possesses single-strand endonuclease activity and double-strand-specific 3'-5' exonuclease activity, which are provided by MRE11A. RAD50 may be required to bind DNA ends and hold them in close proximity. This could facilitate searches for short or long regions of sequence homology in the recombining DNA templates, and may also stimulate the activity of DNA ligases and/or restrict the nuclease activity of MRE11A to prevent nucleolytic degradation past a given point. The complex may also be required for DNA damage signaling via activation of the ATM kinase. In telomeres the MRN complex may modulate t-loop formation.
  • Tissue specificity
    Expressed at very low level in most tissues, except in testis where it is expressed at higher level. Expressed in fibroblasts.
  • Involvement in disease
    Defects in RAD50 are the cause of Nijmegen breakage syndrome-like disorder (NBSLD) [MIM:613078]; also called NBS-like disorder or RAD50 deficiency. NBSLD is a disorder similar to Nijmegen breakage syndrome and characterized by chromosomal instability, radiation sensitivity, microcephaly, growth retardation, short stature and bird-like face. Immunodeficiency is absent.
  • Sequence similarities
    Belongs to the SMC family. RAD50 subfamily.
    Contains 1 zinc-hook domain.
  • Domain
    The zinc-hook, which separates the large intramolecular coiled coil regions, contains 2 Cys residues that coordinate one molecule of zinc with the help of the 2 Cys residues of the zinc-hook of another RAD50 molecule, thereby forming a V-shaped homodimer. The two heads of the homodimer, which constitute the ATP-binding domain, interact with the MRE11A homodimer.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Nucleus. Chromosome > telomere. Localizes to discrete nuclear foci after treatment with genotoxic agents.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA repair protein RAD50 antibody
    • hRAD50 antibody
    • hsRAD50 antibody
    • Mrell antibody
    • NBSLD antibody
    • Rad 50 antibody
    • RAD50 2 antibody
    • rad50 antibody
    • RAD50 homolog (S cerevisiae) antibody
    • RAD50 homolog antibody
    • RAD50 PEN antibody
    • RAD50, S. cerevisiae, homolog of antibody
    • RAD50_HUMAN antibody
    • RAD502 antibody
    • Rad50l antibody
    • Truncated RAD50 protein antibody
    see all

Images

  • Flow Cytometry analysis of U-2 OS (Human bone osteosarcoma epithelial cell) cells labeling Rad50 with purified ab124682 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Rad50 with Purified ab124682 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 . ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/5000 dilution (purified)

    Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
    Lane 2 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 154 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (purified)

    Lane 1 : Human fetal brain lysates
    Lane 2 : Mouse thymus lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 154 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (purified) + Rat testis lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 154 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Unpurified ab124682 staining Rad50 in the U2OS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 5% BSA for 60 minutes. Samples were incubated with undiluted primary antibody  for 2 hours. An undiluted FITC conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Rad50 antibody [EPR3466(2)] (ab124682) at 1/1000 dilution (unpurified)

    Lane 1 : K562 cell lysate
    Lane 2 : MOLT4 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : MCF7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 154 kDa
    Observed band size: 150 kDa (why is the actual band size different from the predicted?)

References

ab124682 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Denaturing (8%)
Sample
Human Cell lysate - whole cell (MEFs, U2OS, HEK 293 and Fibroblasts)
Specification
MEFs, U2OS, HEK 293 and Fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Nov 07 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 03 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HeLa)
Specification
HeLa
Treatment
10 Gy IR
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Apr 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (U2OS)
Specification
U2OS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TritonX100 in PBS
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Nov 19 2012

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

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