Overview

  • Product name
    Anti-Rad51 antibody [51RAD01]
    See all Rad51 primary antibodies
  • Description
    Mouse monoclonal [51RAD01] to Rad51
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, WB, IP, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    BALB/C mice were injected with recombinant Rad51 protein.

  • Positive control
    • Testis

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.05% Sodium azide
    Constituent: 1% BSA
  • Concentration information loading...
  • Purity
    IgG fraction
  • Clonality
    Monoclonal
  • Clone number
    51RAD01
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab1837 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
Flow Cyt 1/100.

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

 

WB Use at an assay dependent concentration. Predicted molecular weight: 37 kDa.
IP Use at an assay dependent concentration.
ICC/IF 1/10. PMID 18663141
  • Application notes
    Is unsuitable for IHC-P.
  • 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

    • Overlay histogram showing HeLa cells stained with ab1837 (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 (ab1837, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was goat anti-mouse DyLight® 488 goat (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 antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Immunofluorescence analysis of wild-type (+/+) and Hormad1-/- (-/-) murine zygotene spermatocytes, staining Rad51 with ab1837.

    References

    This product has been referenced in:
    • Venkatachalam G  et al. Replication stress-induced endogenous DNA damage drives cellular senescence induced by a sub-lethal oxidative stress. Nucleic Acids Res 45:10564-10582 (2017). Read more (PubMed: 28985345) »
    • Ho TT  et al. Autophagy maintains the metabolism and function of young and old stem cells. Nature 543:205-210 (2017). Read more (PubMed: 28241143) »
    See all 14 Publications for this product

    Customer reviews and Q&As

    1-10 of 25 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela)
    Gel Running Conditions
    Non-reduced Denaturing (12%)
    Loading amount
    10 µg
    Specification
    Hela
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Oct 21 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Hela)
    Permeabilization
    Yes - Triton X100
    Specification
    Hela
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 20 2017

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Planarian Cell (whole animal IF)
    Permeabilization
    Yes - tritonX-100
    Specification
    whole animal IF
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jul 20 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
    Sample
    Mouse Tissue sections (mouse testes)
    Specification
    mouse testes
    Permeabilization
    No
    Fixative
    4% PFA

    Abcam user community

    Verified customer

    Submitted May 29 2013

    Answer

    That is no problem at all. I am glad to have been of some help. Please do let me know how you get on.

    Read More

    Answer

    Thank you for your email. I am sorry i was not able to get back to you yesterday.

    I can confirm we have had no other reports of concerns with the lot of the antibody you have been using. I have therefore arranged for a new vial of this lot to be sent to you. If you have similar problems again, please do let me know. You would of course be covered by the Abpromise and would be entitled to a replacement antibody or refund if the problems persist.

    The order number is xxxxx (Purchase order number xxxxx). If you have any problems receiving this please do let me know.

    I hope this replacement vial performs better for you. I look forward to hearing how you get on.

    Until then, I wish you all the best with your research.

    Read More

    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.

    Read More

    Answer

    Thank you for your reply and confirming those details.

    I think it would be worthwhile to try the different blocking conditions I suggested. Sometimes different antibodies can react quite differently using different blocking conditions. This can be clearly seen with the ab9385 where Western blotting using milk produces a much weaker signal than when using BSA. This is illustrated in the following datasheet:

    www.abcam.com/ab9385

    One thing that is a little inconsistent with this theory is that the staining in the leptotene and pachytene especially show more staining in the 1/10 dilution than the 1/2 dilution. But from your next staining we should be able to draw further conclusions.

    May I ask, have you performed any "no-primary" or isotype controls to assess if the secondary antibody is contributing any non-specificity?

    On a different topic, I’d like to bring your attention to a very good offer we are running throughout November. If you order any primary antibody you can receive a RabMab free, whilst stocks last. Simply quote “RABMAB-XBSMG” in your next primary antibody order. More information on this offer can be found from the following link:

    https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    I look forward to hearing how you get on.

    Read More

    Answer

    Thank you for taking time to contact us and report the problems you have encountered in using the Anti-Rad51 antibody [51RAD01] (ab1837). I am sorry to hear that this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

    Having reviewed this case, there are a few points I would suggest optimising in the protocol used. Whilst as you say, the rabbit polyclonal you have developed in your lab looks like it is producing stronger staining, the ab1837 staining localisation loooks identical, only weaker. I would therefore suggest it may be the blocking conditions currently used which may be too strong for this antibody.

    In order to improve this I would suggest trying the following:

    1. I would use either BSA or milk blocking, one at a time to see which performs the best with the anitbody. As a blocking step I would suggest using 3% BSA or Milk in PBST for 1 hour ar room tem.perature In the antibody diluent I would only include 0.1% BSA or milk in PBST to start with. If the background increases, this can always be increased in turn. I would stick with a 1/10 dilution of the primary antibody.

    2. You have mentioned that you have used a goat anti-rabbit Alexafluor antibody for detection. I would check this as the ab1737 is a mouse IgG1 antibody. If you have been using an anti-rabbit secondary antibody you need to change this to an anti-mouse IgG.

    Should the suggestions not improve the results, please do let me know.

    In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

    I hope this information is helpful, and I thank you for your cooperation.

    Read More

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1174833.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    1-10 of 25 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up