Anti-Rad51 antibody [51RAD01] (ab1837)

Reviews (9)Q&A (19)References (16)

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

      see all

    • 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

    • Flow Cytometry - Anti-Rad51 antibody [51RAD01] (ab1837)
      Flow Cytometry - Anti-Rad51 antibody [51RAD01] (ab1837)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [51RAD01] (ab1837)
      Immunocytochemistry/ Immunofluorescence - Anti-Rad51 antibody [51RAD01] (ab1837)Image from Shin YH et al., PLoS Genet. 2010 Nov 4;6(11):e1001190.Fig 5.; doi:10.1371/journal.pgen.1001190November 4, 2010, PLoS Genet 6(11): e1001190.

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

    References

    This product has been referenced in:

    • Bodo S  et al. Single-dose radiotherapy disables tumor cell homologous recombination via ischemia/reperfusion injury. J Clin Invest 129:786-801 (2019). Read more (PubMed: 30480549) »
    • 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 16 Publications for this product

    Publishing research using ab1837? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 19 Q&A

    Question

    Dear xxxx,

    Thank you for your email. I hope the experiment will work better with this new vial.

    I will let you know about the outcome in the coming weeks.

    Thank you very much for your prompt replies and for your helpfulness.

    Best regards,

    Read More
    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.

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    Question

    Dear xxxx,

    I appreciate your helpfulness in trying to solve this problem. I was wondering whether some other customer gave you negative feedback as regards the ab1837 antibody from this batch. If this is not the case, it is more likely that something was wrong only for the aliquote we received. We would like to get a new vial of the same ab1837 antibody and give it one more try. In case we run into the same problem, then we would consider the option of a refund or of buying a different antibody.

    You asked some details to track our order: I have the "purchase number" xxxxx

    In case you need further details about the ordering procedure, please do not hesitate to contact me again.

    Best regards,

    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.

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    Question

    Dear xxxx,

    As I told you in my previous mail, I repeated the immunostaining following your suggestion of testing different blocking conditions and changing the antibody diluent to a less concentrated BSA solution.

    Unfortunately this only increased my background but the quality of the staining did not increase. It is still not possible to detect RAD51 foci with the ab1837 that we bought. Our secondary antibody is not contributing either to the background staining.

    I think it is of interest for you to know that we tested also ab133534 (rabbit monoclonal) on our preparations in the usual conditions (the ones I described in my first enquiry): this antibody works quite well (1:250) although it gives a little bit too much background compared to our homemade rabbit polyclonal.

    Taking all this together, I would guess that there is really something wrong with the ab1837 that we received.

    You mentioned in your previous reply that you could provide a replacement in case the antibody would still not work. If that is still an option, we would like to proceed that way, also because there are published data showing a quite good staining of RAD51 with ab1837 in testis spread preparations.

    I am looking forward to hearing from you soon

    Kind regards,

    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.

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    Question

    Dear xxxx,

    First of all, I want to thank you for replying so quickly to my enquiry.

    I am going to try what you suggest about the blocking conditions, although we apply the conditions I described in my previous message for immunostainings with all the antibodies we use (including some more Abcam mouse monoclonals, such as anti-DMC1).

    Anyways, I do not see such clear foci pattern in the nuclei I analyzed so far, because the background spots are too similar to what I should consider foci. In such situation I could not objectively discriminate a focus from a random green dot in the nucleus and it would not be fair analysis if I would consider a dot as a focus just because it is located on the axes of the synaptonemal complex.

    I also want to reassure you about the secondary antibody: I made a mistake while typing the message. Since it was a combined immunostaining with a rabbit primary antibody for the synaptonemal complex, I got confused. However, I used a goat anti mouse Alexafluor 488 to detect your ab1837.

    I will let you know whether things work out fine or not as soon as possible.

    Once again, thank you very much for your assistance and helpfulness.

    Best regards,

    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

    Question

    Product code: 1837
    Lot number: GR98318-1

    Inquiry: I used the antibody for IF staining of RAD51 in mouse spermatocytes spread preparations fixed in 1% PFA (+0.15% Triton X-100). Blocking 0.5%BSA 0.5%Milk in 1X PBS Antibody was diluted 1/10 and 1/2 in 10%BSA. Incubation was performed overnight at 4 degrees or at RT in a humid chamber. Detection was performed with goat anti rabbit Alexa fluor 488 (1/1000) for 2hrs RT in a humidified atmosphere. Washings between the steps were performed with 1X PBS. The immuno never worked. The same preparations have been stained for RAD51 with a homemade rabbit polyclonal antibody and it worked perfectly. Please find some representative pictures in the attachment. Since this antibody has been used on testis spreads by another research group which showed pictures in a published paper, we really expected this antibody to work.

    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

    Question

    I have tried to usethe anti-Rad51 antibodyusing methanol fixation and bloking for 2 h with 5% BSA as you suggested. However, the antibody did not work.
    Is it possible to have some more anti-Rad51 antibody (ab1837) as this product was purchased less than six months ago?
    Best Wishes,

    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

    Question

    Mycolleague blocked his cells with 4%BSA overnight and it did not work for him as well. I haven't had any problem before usingparaformaldehyde as a fixing agent.In fact the protocol did work forDNAPKcs antibody.
    Could you please let me know the exposure timeand concentration of methanol? Also, if I keep the samples in the fridge for few days before antibody staining, should I leave them in the fixing agent or simply PBS?
    Best Wishes,

    Read More
    Answer

    Thank you for contacting us.

    Ice-cold methanol for 5-10 minutes would be ideal.

    I would suggest not storing the samples however if it is unavoidable then storing cells after fixation on chamber slides would be the option to go for.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    Question

    Attached the completed form and a picture.
    Best Wishes,

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    We haven't received any other complaint for this lot so I can reassure you that the quality of this antibody is absolutely fine. I have read the details you have kindly provided and presuming that the protocol troubleshooting will help. I have following suggestions;

    - Methanol fixation: sometime antibodies works better with methanol fixation rather than PFA so you may try this.
    - Try blocking with 5% BSA for 1-2 hours.

    I hope these suggestion would help to improve results. If in case the results do not improve please contact for replacement product.

    Read More

    Question

    Problem with antibody ab1837.

    Read More
    Answer

    Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, I amattaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

    Read More

    Question

    BATCH NUMBER 169563 ORDER NUMBER 152012 DESCRIPTION OF THE PROBLEM Immediately after arrival, worked fine for Western. A week later the aliquot at 4 deg tested for IF but gave no signal. A couple of days ago we tested fresh aliquot from - 20 deg in Western but obtained no signal. The blots were also stained with two other antibodies and worked fine. Exactly the same problem occurred twice with this antibody bought by my collaborators in Sofia, Bulgaria, within the last 6 months. SAMPLE HeLa and PC3 cell whole cell extracts for Western. HeLa and PC3 cells irradiated with X-rays tested for IF. PRIMARY ANTIBODY 1:1000 dilution in TBS, overnight incubation at 4 deg. Washed in TBST, 5 times for 5 min. DETECTION METHOD Licor Odyssey infrared scanner. ANTIBODY STORAGE CONDITIONS 1 aliquot stored at 4 deg ALiquots at -20 deg SAMPLE PREPARATION Cells resuspended in CSK buffer with leupeptin, pepstatin, DTT and complete protease inhibitor (Roche). Standard SDS gel loading buffer, heated at 100 deg. AMOUNT OF PROTEIN LOADED At least 20 ug loaded per lane. ELECTROPHORESIS/GEL CONDITIONS Reducing SDS PAGE, 12.5%. TRANSFER AND BLOCKING CONDITIONS Semi-dry transfer, 25V, 45 min. Blocking: 5% dry skimmed milk in TBS, 1h room temp. SECONDARY ANTIBODY AlexaFluor 680 conjugated goat anti-mouse IGG, MOlecular Probes. 1:2000 dilution. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? There is no point to alter steps as the protocol we used at first gave good result. Moreover, we did not get immunofluorescence signal with this antibody. Attempts to use lower dilutions were carried out by my collaborators, who experienced similar problems with this antibody in Bulgaria, were unsuccessul. ADDITIONAL NOTES We have used this antibody extensively in the past. In fact, you are citing two of our papers in the information provided for this antibody. Problems started after you started shipping a diluted version of the antibody.

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    Answer

    I'm very sorry to hear you have experienced problems with ab1837 and thank you for providing your protocol details. I have forwarded your complaint to the source of this antibody for their feedback and take your comments extremely seriously. Would you like a replacement vial or would you prefer a refund? I look forward to hearing from you and apologise for this problem,

    Read More

    1-10 of 19 Q&A

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