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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

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

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.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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