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
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • 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

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    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

  • 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: 7.40
    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

Applications

Our Abpromise guarantee covers the use of ab133534 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
WB 1/10000 - 1/50000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/1000.

For unpurified use at 1/100 - 1/250.

Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/10 - 1/100.

Target

  • Function
    Plays an important role in homologous strand exchange, a key step in DNA repair through homologous recombination. Binds to single and double-stranded DNA and exhibits DNA-dependent ATPase activity. Catalyzes the recognition of homology and strand exchange between homologous DNA partners to form a joint molecule between a processed DNA break and the repair template. Binds to single-stranded DNA in an ATP-dependent manner to form nucleoprotein filaments which are essential for the homology search and strand exchange (PubMed:26681308). Part of a PALB2-scaffolded HR complex containing BRCA2 and RAD51C and which is thought to play a role in DNA repair by HR. Plays a role in regulating mitochondrial DNA copy number under conditions of oxidative stress in the presence of RAD51C and XRCC3.
  • Tissue specificity
    Highly expressed in testis and thymus, followed by small intestine, placenta, colon, pancreas and ovary. Weakly expressed in breast.
  • Involvement in disease
    Breast cancer
    Mirror movements 2
    Defects in RAD51 are found in a patient with microcephaly, mental retardation without bone marrow failure and pediatric cancers.
  • Sequence similarities
    Belongs to the RecA family. RAD51 subfamily.
    Contains 1 HhH domain.
  • Domain
    The nuclear localization may reside in the C-terminus (between 259 and 339 AA).
  • Post-translational
    modifications
    Ubiquitinated by the SCF(FBXO18) E3 ubiquitin ligase complex, regulating RAD51 subcellular location and preventing its association with DNA.
    Phosphorylated. Phosphorylation of Thr-309 by CHEK1 may enhance association with chromatin at sites of DNA damage and promote DNA repair by homologous recombination. Phosphorylation by ABL1 inhibits function.
  • Cellular localization
    Nucleus. Cytoplasm. Cytoplasm, perinuclear region. Mitochondrion matrix. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Colocalizes with RAD51AP1 and RPA2 to multiple nuclear foci upon induction of DNA damage. DNA damage induces an increase in nuclear levels. Together with FIGNL1, redistributed in discrete nuclear DNA damage-induced foci after ionizing radiation (IR) or camptothecin (CPT) treatment. Accumulated at sites of DNA damage in a SPIDR-dependent manner.
  • Information by UniProt
  • Database links
  • Alternative names
    • BRCA1/BRCA2 containing complex, subunit 5 antibody
    • BRCC 5 antibody
    • BRCC5 antibody
    • DNA repair protein RAD51 homolog 1 antibody
    • DNA repair protein rhp51 antibody
    • FANCR antibody
    • hRAD51 antibody
    • HsRAD51 antibody
    • HsT16930 antibody
    • MRMV2 antibody
    • Rad 51 antibody
    • RAD51 antibody
    • RAD51 homolog (RecA homolog, E. coli) (S. cerevisiae) antibody
    • RAD51 homolog A antibody
    • RAD51 homolog antibody
    • RAD51 recombinase antibody
    • RAD51, S. cerevisiae, homolog of antibody
    • RAD51_HUMAN antibody
    • RAD51A antibody
    • RECA antibody
    • RecA like protein antibody
    • RecA, E. coli, homolog of antibody
    • Recombination protein A antibody
    see all

Images

  • 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).

  • 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) 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 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.

  • 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.

  • 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.

  • 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.

  • 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) analysis of human testis tissue labelling Rad51 with unpurified ab133534 at a dilution of 1/100.

  • 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Zhong Q  et al. Cyclin D1 silencing impairs DNA double strand break repair, sensitizes BRCA1 wildtype ovarian cancer cells to olaparib. Gynecol Oncol 152:157-165 (2019). Read more (PubMed: 30414739) »
  • Luo J  et al. MicroRNA-146a-5p Enhances Radiosensitivity in Hepatocellular Carcinoma through Replication Protein A3 Induced Activation of the DNA Repair Pathway. Am J Physiol Cell Physiol N/A:N/A (2018). Read more (PubMed: 30462539) »
See all 14 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Glioblastoma cells U87)
Permeabilization
Yes - PBST 0.3%
Specification
Glioblastoma cells U87
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Mr. Matthias Dedobbeleer

Verified customer

Submitted Jun 01 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (Bovine ovaries (cortex))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate
Permeabilization
No
Specification
Bovine ovaries (cortex)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 23°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 27 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Casein as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Sample
Human Tissue sections (testis)
Specification
testis
Permeabilization
No
Fixative
Paraformaldehyde

Dr. Henry King

Verified customer

Submitted Dec 24 2013

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (Glioblastoma tumour stem cells)
Specification
Glioblastoma tumour stem cells
Blocking step
Odyssey Blocking Buffer as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Dr. Henry King

Verified customer

Submitted Nov 20 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Schmidtea mediterranea Cell lysate - whole cell (Whole body)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (15)
Loading amount
30 µg
Specification
Whole body
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 14 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Tissue sections (Brain tumour samples)
Specification
Brain tumour samples
Permeabilization
No
Fixative
Methanol

Dr. Henry King

Verified customer

Submitted Aug 20 2013

Answer

Thank you for your reply and for letting me know how you have been getting on.

I am sorry that my suggestions have not improved the results observed. As you say, especially with the positive results with the rabbit monoclonal ab133534, it may be that the vial you received may have lost its activity. I am sorry for the inconvenience this has caused you.

If you would like I can provide you with a free vial of ab1837 however, we only have the same lot you have been trying in stock. Alternatively I can provide you with a different antibody altogether such as the chicken polyclonal ab63802 or the rabbit polyclonal ab137323. Or if you would prefer, I could provide you with an alternative antibody against any target of your choice (as you already have ab133534 which seems to be working for you) or a credit note or refund.

In order to arrange for any of these I would need confirmation of the order number on which ab1837 was received. Could you provide this, or if you do not have this information, the approximate delivery date and the delivery address used?

Many thanks for your cooperation. I look forward to hearing how you would like to proceed.

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