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    rad50-antibody-epr34662-ab124682.pdf

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Epigenetics and Nuclear Signaling Chromosome Structure Telomeres
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RecombinantRabMAb

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

  • Datasheet
Reviews (5) Submit a question References (5)

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Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Flow Cytometry (Intracellular) - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
  • Anti-Rad50 antibody [EPR3466(2)] (ab124682)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3466(2)] to Rad50
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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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 (Intra), ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3466(2)
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Chromosome Structure
    • Telomeres
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Non Homol. End Joining
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • Homologous Recomb.

Associated products

  • Alternative Versions

    • Anti-Rad50 antibody [EPR3466(2)] - BSA and Azide free (ab239993)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • K-562 nuclear extract lysate (ab14851)
    • MCF7 nuclear extract lysate (ab14860)

Applications

The Abpromise guarantee

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 (Intra)
1/80.
ICC/IF (3)
1/100.
WB (2)
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 154 kDa).
Notes
Flow Cyt (Intra)
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.
  • Target information above from: UniProt accession Q92878 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10111 Human
    • Entrez Gene: 19360 Mouse
    • Entrez Gene: 64012 Rat
    • Omim: 604040 Human
    • SwissProt: Q92878 Human
    • SwissProt: P70388 Mouse
    • SwissProt: Q9JIL8 Rat
    • Unigene: 633509 Human
    • Unigene: 4888 Mouse
    • Unigene: 51136 Rat
    see all
  • 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

  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    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

  • Flow Cytometry (Intracellular) - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Flow Cytometry (Intracellular) - Anti-Rad50 antibody [EPR3466(2)] (ab124682)

    Intracellular Flow Cytometry analysis of U-2 OS (Human bone osteosarcoma epithelial cell) cells labeling Rad50 with purified ab124682 at 1/80 dilution (10µg/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 - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)

    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.

  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    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

  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Immunocytochemistry/ Immunofluorescence - Anti-Rad50 antibody [EPR3466(2)] (ab124682)This image is courtesy of an anonymous Abreview

    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

  • Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    Western blot - Anti-Rad50 antibody [EPR3466(2)] (ab124682)
    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?

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

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (5)

Publishing research using ab124682? Please let us know so that we can cite the reference in this datasheet.

ab124682 has been referenced in 5 publications.

  • Tang D  et al. MYC/NBS1-Mediated DNA Damage Response is Involved in the Inhibitory Effect of Hydroxysafflor Yellow A on Glioma Cells. Drug Des Devel Ther 15:1749-1763 (2021). PubMed: 33953544
  • Ren S  et al. ATM-mediated DNA double-strand break response facilitated oncolytic Newcastle disease virus replication and promoted syncytium formation in tumor cells. PLoS Pathog 16:e1008514 (2020). PubMed: 32479542
  • Legrand JMD  et al. DDX5 plays essential transcriptional and post-transcriptional roles in the maintenance and function of spermatogonia. Nat Commun 10:2278 (2019). PubMed: 31123254
  • Chen Y  et al. EGFR tyrosine kinase inhibitor HS-10182 increases radiation sensitivity in non-small cell lung cancers with EGFR T790M mutation. Cancer Biol Med 15:39-51 (2018). PubMed: 29545967
  • Sun C  et al. BRD4 Inhibition Is Synthetic Lethal with PARP Inhibitors through the Induction of Homologous Recombination Deficiency. Cancer Cell 33:401-416.e8 (2018). PubMed: 29533782

Customer reviews and Q&As

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1-5 of 5 Abreviews

Western blot abreview for Anti-Rad50 antibody [EPR3466(2)]

Good
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Nov 07 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad50 antibody [EPR3466(2)]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Nov 03 2014

Western blot abreview for Anti-Rad50 antibody [EPR3466(2)]

Excellent
Abreviews
Abreviews
abreview image
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%
Read More

Abcam user community

Verified customer

Submitted Apr 25 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad50 antibody [EPR3466(2)]

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Apr 23 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad50 antibody [EPR3466(2)]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% TritonX100 in PBS
Read More

DR. Kirk Mcmanus

Verified customer

Submitted Nov 19 2012

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