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    rad50-antibody-13b32c6-ab89.pdf

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Epigenetics and Nuclear Signaling Chromosome Structure Telomeres
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Anti-Rad50 antibody [13B3/2C6] (ab89)

  • Datasheet
  • SDS
Reviews (6)Q&A (3)References (40)

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Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)
  • Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad50 antibody [13B3/2C6] (ab89)
  • Flow Cytometry - Anti-Rad50 antibody [13B3/2C6] (ab89)

Key features and details

  • Mouse monoclonal [13B3/2C6] to Rad50
  • Suitable for: Flow Cyt, IHC-P, WB
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-Rad50 antibody [13B3/2C6]
    See all Rad50 primary antibodies
  • Description

    Mouse monoclonal [13B3/2C6] to Rad50
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Fusion protein containing the complete coding region (amino acids 1-425) of RAD50 expressed in E.coli.

  • Positive control

    • Raji cell lysate HEK293 cell lysate
  • General notes

    This product was changed from ascites to tissue culture supernatant on 01/02/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    Clone 2C6 recognizes Rad50, a 153kDa protein involved in DNA double strand break repair. Rad50 is associated with Mre11 and p95 (Nibrin) to form a multiprotein complex involved in the double strand break repair process.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.40
    Constituent: 100% PBS
  • Concentration information loading...
  • Purity

    Affinity purified
  • Purification notes

    Affinity purified by Protein G.
  • Clonality

    Monoclonal
  • Clone number

    13B3/2C6
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Light chain type

    kappa
  • 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

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab89 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P
1/100 - 1/1000.
WB (5)
1/500 - 1/3000. Detects a band of approximately 150 kDa (predicted molecular weight: 146 kDa).
Notes
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P
1/100 - 1/1000.
WB
1/500 - 1/3000. Detects a band of approximately 150 kDa (predicted molecular weight: 146 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
    • Omim: 604040 Human
    • SwissProt: Q92878 Human
    • Unigene: 633509 Human
    • 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 [13B3/2C6] (ab89)
      Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)

      This picture was kindly supplied as part of the reviews for ab214 and for ab89 submitted by Anya Polischouk.

      The bands represent nuclear (N) and cytoplasmic (C) extract from human lung cancer cells (U1810).

      This picture was kindly supplied as part of the reviews for ab214 and for ab89 submitted by Anya Polischouk.

      The bands represent nuclear (N) and cytoplasmic (C) extract from human lung cancer cells (U1810).

      This image was generated using the ascites version of the product.

    • Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)
      Western blot - Anti-Rad50 antibody [13B3/2C6] (ab89)
      Anti-Rad50 antibody [13B3/2C6] (ab89) at 1/1000 dilution + 50 ug Human HEK293 total cell lysate

      Secondary
      HRP conjugated Donkey Anti-Mouse IgG

      Developed using the ECL technique.

      Performed under non-reducing conditions.

      Predicted band size: 146 kDa
      Observed band size: 160 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 300 kDa (possible non-specific binding)


      Exposure time: 30 seconds


      This image is courtesy of an Abreview submitted by Philippe Szankasi on 24 February 2006.

      This image was generated using the ascites version of the product.

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad50 antibody [13B3/2C6] (ab89)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad50 antibody [13B3/2C6] (ab89)

      ab89 (1µg/ml) staining RAD50 in human placenta, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

      This image was generated using the ascites version of the product.

    • Flow Cytometry - Anti-Rad50 antibody [13B3/2C6] (ab89)
      Flow Cytometry - Anti-Rad50 antibody [13B3/2C6] (ab89)

      Overlay histogram showing HeLa cells stained with ab89 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab89, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

      This image was generated using the ascites version of the product.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (40)

    Publishing research using ab89? Please let us know so that we can cite the reference in this datasheet.

    ab89 has been referenced in 40 publications.

    • Meng X  et al. DNA damage repair alterations modulate M2 polarization of microglia to remodel the tumor microenvironment via the p53-mediated MDK expression in glioma. EBioMedicine 41:185-199 (2019). PubMed: 30773478
    • Hwang SY  et al. CTCF cooperates with CtIP to drive homologous recombination repair of double-strand breaks. Nucleic Acids Res 47:9160-9179 (2019). PubMed: 31340001
    • Babeu JP  et al. Quantitative Proteomics Identifies DNA Repair as a Novel Biological Function for Hepatocyte Nuclear Factor 4a in Colorectal Cancer Cells. Cancers (Basel) 11:N/A (2019). PubMed: 31060309
    • Chang EY  et al. MRE11-RAD50-NBS1 promotes Fanconi Anemia R-loop suppression at transcription-replication conflicts. Nat Commun 10:4265 (2019). PubMed: 31537797
    • Ho V  et al. Overexpression of the MRE11-RAD50-NBS1 (MRN) complex in rectal cancer correlates with poor response to neoadjuvant radiotherapy and prognosis. BMC Cancer 18:869 (2018). PubMed: 30176843
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (WM266 human melanoma)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    20 µg
    Specification
    WM266 human melanoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 23 2018

    Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (293T and MEF)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    293T and MEF
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 22 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad50 antibody [13B3/2C6]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Vascular smooth muscle cell)
    Permeabilization
    Yes - NP40
    Specification
    Vascular smooth muscle cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 12 2018

    Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SW480 cells)
    Loading amount
    25 µg
    Specification
    SW480 cells
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 05 2013

    Question

    I am looking for positive and negative controls for this antibody.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 22 2007

    Answer

    Thank you for your enquiry. I was looking at the Swiss Prot entry for the protein that this antibody is specific too, and the tissue specificity is listed as: Expressed at very low level in most tissues, except in testis where it is expressed at higher level. Expressed in fibroblasts. That being said, I think it might be difficult to find a negative control, perhaps you would have some luck in literature. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Jan 24 2007

    Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (lung and foreskin fibroblast cells)
    Specification
    lung and foreskin fibroblast cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 02 2006

    Western blot abreview for Anti-Rad50 antibody [13B3/2C6]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5%
    Read More

    Dr. Philippe Szankasi

    Verified customer

    Submitted Feb 24 2006

    Question

    I have a customer who wishes to know whether the epitope for this product has been mapped. Is the epitope located in/around the N-terminus? Thank you for your help.

    Read More

    Abcam community

    Verified customer

    Asked on Jan 06 2005

    Answer

    Thank you for inquiring about our RAD50 monoclonal antibody (clone 13B3). The exact epitope that this antibody recognizes has not been determined. However, it is quite possible that the epitope is near or around the amino terminus of RAD50 as the immunogen used for producing this monoclonal comprised of amino acids 1-425 of the protein. I apologise for not being able to offer nay further information regarding the epitope, if you have any further questions then please get back in touch with us.

    Read More

    Abcam Scientific Support

    Answered on Jan 06 2005

    Question

    In order to use this antibody for immunostaining, can you suggest me some informations about the dilution conditions? Thank you.carmela Ciotta

    Read More

    Abcam community

    Verified customer

    Asked on Nov 22 2000

    Answer

    Dear Carmela, Have Genetex replied to you on this one yet? Abcam

    Read More

    Abcam Scientific Support

    Answered on Dec 13 2000

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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