Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Rad51 antibody [EPR4030(3)] (ab133534)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4030(3)] to Rad51
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Rad51 antibody [EPR4030(3)]
    See all Rad51 primary antibodies

  • Description

    Rabbit monoclonal [EPR4030(3)] to Rad51

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Rad51 aa 1-100. The exact sequence is proprietary.
    Database link: Q06609
    (Peptide available as ab176353)

  • Positive control

    • WB: C6, NIH/3T3, 293T, Jurkat, HeLa, and K562 cell lysates and mouse spleen tissue lysate. IHC-P: Human cervix carcinoma and testis tissues. ICC/IF: Jurkat cells. Flow Cyt: HeLa cells. IP: HEK293 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. Stable for 12 months at -20°C.

  • Dissociation constant (KD)

    KD = 9.20 x 10 -11 M
    Learn more about KD

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR4030(3)

  • Isotype

    IgG

  • Research areas

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat (human T cell leukemia cell line from peripheral blood) cells labelling Rad51 with purified ab133534 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/1000) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    All lanes : Anti-Rad51 antibody [EPR4030(3)] (ab133534) at 1/10000 dilution (purified)

    Lane 1 : C6 (rat glial tumor cell line) whole cell lysate
    Lane 2 : Mouse spleen tissue lysate
    Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 37 kDa
    Observed band size: 37 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling Rad51 with purified ab133534 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Flow Cytometry - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Flow Cytometry - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling Rad51 with purified ab133534 at 1/350 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunoprecipitation - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Immunoprecipitation - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    ab133534 (purified) at 1/100 immunoprecipitating Rad51 in HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate.

    Lane 1 (input): HEK-293 whole cell lysate (10µg)

    Lane 2 (+): ab133534 + HEK-293 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133534 in HEK-293 whole cell lysate.

    For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [EPR4030(3)] (ab133534) Gao et al Nat Commun. 2018; 9: 3925. Published online 2018 Sep 25. doi: 10.1038/s41467-018-06407-7

    Immunocytochemical analysis of DT40 cells (WT and the indicated KOs) labeling Rad51 using ab133534 at 1/200 dilution.

    From Figure 5d of gao et al.

    Gao et al Nat Commun. 2018; 9: 3925. Published online 2018 Sep 25. doi: 10.1038/s41467-018-06407-7

    Reproduced under the Creative Commons Licence http://creativecommons.org/licenses/by/4.0/.

  • Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Anti-Rad51 antibody [EPR4030(3)] (ab133534) at 1/10000 dilution (purified) + K562 (human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 37 kDa
    Observed band size: 37 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    All lanes : Anti-Rad51 antibody [EPR4030(3)] (ab133534) at 1/20000 dilution (purified)

    Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 37 kDa
    Observed band size: 37 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Western blot - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    All lanes : Anti-Rad51 antibody [EPR4030(3)] (ab133534) at 1/10000 dilution (unpurified)

    Lane 1 : C6 (rat glial tumor cell line) cell lysate
    Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 5 : K562 (human chronic myelogenous leukemia cell line from bone marrow ) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 37 kDa
    Observed band size: 37 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Rad51 with unpurified ab133534 at a dilution of 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Flow Cytometry - Anti-Rad51 antibody [EPR4030(3)] (ab133534)

    Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with unpurified ab133534 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (unpurified ab133534, 1/1000 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1 μg/1x106 cells) used under the same conditions. Unlabeled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • OI-RD Scanning - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    OI-RD Scanning - Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-Rad51 antibody [EPR4030(3)] (ab133534)
    Anti-Rad51 antibody [EPR4030(3)] (ab133534)

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