Overview

  • Product name
    Anti-Rad51 antibody - ChIP Grade
    See all Rad51 primary antibodies
  • Description
    Rabbit polyclonal to Rad51 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, ChIP, ICC/IF, Dot blot, Indirect ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Chicken, Human
    Predicted to work with: Rabbit, Cow, Dog, Xenopus laevis, Chinese hamster
  • Immunogen

    Recombinant full length protein corresponding to Human Rad51 aa 1-339.
    Sequence:

    MAMQMQLEANADTSVEEESFGPQPISRLEQCGINANDVKKLEEAGFHTVE AVAYAPKKELINIKGISEAKADKILAEAAKLVPMGFTTATEFHQRRSEII QITTGSKELDKLLQGGIETGSITEMFGEFRTGKTQICHTLAVTCQLPIDR GGGEGKAMYIDTEGTFRPERLLAVAERYGLSGSDVLDNVAYARAFNTDHQ TQLLYQASAMMVESRYALLIVDSATALYRTDYSGRGELSARQMHLARFLR MLLRLADEFGVAVVITNQVVAQVDGAAMFAADPKKPIGGNIIAHASTTRL YLRKGRGETRICKIYDSPCLPEAEAMFAINADGVGDAKD


    Database link: Q06609

  • Positive control
    • WB: MCF7, NIH3T3, CHO and HeLa whole cell lysate (ab150035); Xenopus laevis egg lysate. IP: HeLa whole cell lysate (ab150035). ICC/IF: GM0637 cells. IHC-P: Mouse testis tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab176458 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 1/1000 - 1/10000. Predicted molecular weight: 36 kDa.
IP 1/100 - 1/1000.
ChIP Use at an assay dependent concentration.
ICC/IF 1/1000 - 1/10000.
Dot blot 1/1000 - 1/5000.
Indirect ELISA 1/2000 - 1/5000.
IHC-P Use at an assay dependent concentration.

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

  • All lanes : Anti-Rad51 antibody - ChIP Grade (ab176458) at 1/1000 dilution

    Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
    Lane 2 : NIH3T3 (Mouse embryo fibroblast cell line) cell lysate
    Lane 3 : CHO (Chinese hamster ovary cell line) cell lysate
    Lane 4 : Xenopus laevis egg

    Lysates/proteins at 40 µg per lane.

    Predicted band size: 36 kDa

  • Immunofluorescence detection of Rad51 foci formation after X-ray irradiation in GM0637 cells with ab176458 at 1/10000 dilution (left panels) and 1/1000 dilution (right panels).  The secondary antibody, anti-rabbit Alexa 488 was used at 1/10000 dilution.

     

    Cells were irradiated by X-rays at 2 Gy, grown for 1 hr, fixed with 4% paraformaldehyde in 1x PBS for 10 min, washed 3 times with PBS for 3 min, permealized by treatment with 0.5% Triton for 5 min, washed 3 times with PBS for 3 min, incubated with ab176458 for 30 min at 37°C, washed 3 times with PBS for 3 min, incubated with secondary antibody for 30 min at 37°C, washed 3 times with PBS for 3 min, stained with Hoechst for 1 min and mounted.

     

    The pictures were by courtesy of Prof. S. Tashiro and Dr. K. Kono at Hiroshima University.

  • Anti-Rad51 antibody - ChIP Grade (ab176458) at 1/1000 dilution + Crude HeLa cell extracts at 10 µg

    Secondary
    Goat anti-Rabbit IgG conjugated to HRP at 1/20000 dilution

    Predicted band size: 36 kDa

  • ab176458 (20 µg) was incubated with 20 μg of HeLa cell extract, and precipitated with 20 μg of proteinA-beads. The sample was dissociated from the precipitate by heating in SDS-sample buffer and analyzed by western blotting with anti-Rad51 antiserum (chicken, ab63802) at 1/1000 dilution. As secondary antibody, anti-chicken IgG antibody (rabbit) was used at 1/10000 dilution.

  • Immunohistological staining of Rad51 protein in mouse testis using ab176458. A section of formalin fixed and paraffin embedded mouse testis was treated with ab176458 at 1/100 dilution after deparaffization and antigen retrieval. The secondary antibody, Alexa Fluor® 647conjugated anti-rabbit IgG was used at 1/1,000 dilution (top left). The sample was counter-stained with DAPI (top right) and the merged image is shown (bottom left). The white light image of the same region is shown on the bottom right.

References

This product has been referenced in:
  • Chakraborty S  et al. SMARCAD1 Phosphorylation and Ubiquitination Are Required for Resection during DNA Double-Strand Break Repair. iScience 2:123-135 (2018). Read more (PubMed: 29888761) »
  • Willis NA  et al. Rad51 recruitment and exclusion of non-homologous end joining during homologous recombination at a Tus/Ter mammalian replication fork barrier. PLoS Genet 14:e1007486 (2018). Read more (PubMed: 30024881) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (11)
Sample
Mouse Cell lysate - whole cell (Mouse Pancreatic cancer)
Specification
Mouse Pancreatic cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 27 2014

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (13%)
Sample
Human Tissue lysate - whole (U2OS osteosarcoma)
Specification
U2OS osteosarcoma
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 14 2014

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