Anti-Rad51 antibody [14B4] (ab213)

Reviews (9)Q&A (11)References (65)

Overview

  • Product name

    Anti-Rad51 antibody [14B4]
    See all Rad51 primary antibodies

  • Description

    Mouse monoclonal [14B4] to Rad51

  • Host species

    Mouse

  • Specificity

    Specific for human Rad51.

  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein corresponding to Human Rad51 aa 1-338.
    Database link: Q06609

  • Positive control

    • WB: RAD51 shRNA transfected (+) 293T whole cell extracts, Jurkat, Raji and NCI-H929 whole cell lysate. IHC-P: BT483 xenoograft tissue, human colon esophagus tissue.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 17th September 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • Storage buffer

    pH: 7.40
    Constituent: PBS
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Monoclonal

  • Clone number

    14B4

  • Myeloma

    NS1

  • Isotype

    IgG2b

  • Light chain type

    kappa

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab213 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
IHC-P 1/100 - 1/1000.
WB 1/500 - 1/3000. Predicted molecular weight: 37 kDa.

Please see WB protocol in image legend. 
IP Use at an assay dependent concentration.

Assay dependent
  • Application notes
    Is unsuitable for ICC/IF.

    • 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

      • Western blot - Anti-Rad51 antibody [14B4] (ab213)
        Western blot - Anti-Rad51 antibody [14B4] (ab213)
        All lanes : Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution

        Lane 1 : Jurkat whole cell lysate
        Lane 2 : Raji whole cell lysate
        Lane 3 : NCI-H929 whole cell lysate

        Lysates/proteins at 30 µg per lane.

        Secondary
        All lanes : HRP-conjugated anti-mouse IgG antibody at 1/10000 dilution

        Developed using the ECL technique.

        Predicted band size: 37 kDa



        10% SDS-PAGE

        Running condition: 80V, 15min; 140V, 40 min

        Transfer condition: Semi-dry, 18 V, 60 min (Nitrocellulose membrane)

        Blocking condition: 5% non-fat milk in TBST, RT, 60 min.

        Primary antibody incubation: 4°C overnight.

        Washing condition: 5 ml TBST, 4 x 5 min.

        Chemiluminescent HRP Substrate.

         

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

        Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Rad51 with ab213 at 1/100 dilution at 4°C overnight. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H2O2, RT, 30minutes. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Secondary antibody: ABC HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 mins. Chromogen system: DAB.

      • Western blot - Anti-Rad51 antibody [14B4] (ab213)
        Western blot - Anti-Rad51 antibody [14B4] (ab213)
        All lanes : Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution

        Lane 1 : Non-transfected 293T (-) whole cell extracts
        Lane 2 : RAD51 shRNA transfected (+) 293T whole cell extracts

        Lysates/proteins at 30 µg per lane.

        Secondary
        All lanes : HRP-conjugated anti-mouse IgG antibody

        Developed using the ECL technique.

        Predicted band size: 37 kDa



        10% SDS-PAGE

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

        Immunohistochemical analysis of paraffin-embedded human esophagus tissue labeling Rad51 with ab213 at 1/100 dilution at 4°C overnight. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Endogenous peroxidase blocking: 3% H2O2, RT, 30minutes. Blocking: 1.5% goat serum (dilute goat serum by 1xPBS), RT, 30 min. Secondary antibody: ABC HRP Kit (Mouse IgG), 1:200, RT, 30min. Washing: PBS, 2 x 5 mins. Chromogen system: DAB.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)

        Immunohistochemical analysis of paraffin-embedded BT483 xenoograft tissue labeling RAD51 with ab213 at 1/200 dilution. Nuclear staining is observed. Antigen Retrieval: EDTA based, pH 8.0 buffer, 15min.

      • Western blot - Anti-Rad51 antibody [14B4] (ab213)
        Western blot - Anti-Rad51 antibody [14B4] (ab213)Image courtesy of an anonymous Abreview.
        Anti-Rad51 antibody [14B4] (ab213) at 1/500 dilution + lysate prepared from murine NIH3T3 cells at 20 µg

        Secondary
        HRP conjugated anti-mouse IgG at 1/5000 dilution

        Developed using the ECL technique.

        Predicted band size: 37 kDa
        Observed band size: 42,45 kDa
        why is the actual band size different from the predicted?


        Exposure time: 5 minutes
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad51 antibody [14B4] (ab213)This image is courtesy of an anonymous Abreview

        ab213 staining Rad51 in Human Lovo xenograft tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 20% serum for 20 minutes at room temperature; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/100 in TBST) for 1 hour. An undiluted HRP-conjugated Donkey anti-mouse IgG polyclonal was used as the secondary antibody.

        See Abreview

      References

      This product has been referenced in:

      • Horikoshi N  et al. Pre-existing H4K16ac levels in euchromatin drive DNA repair by homologous recombination in S-phase. Commun Biol 2:253 (2019). Read more (PubMed: 31286070) »
      • Silva B  et al. FANCM limits ALT activity by restricting telomeric replication stress induced by deregulated BLM and R-loops. Nat Commun 10:2253 (2019). Read more (PubMed: 31138795) »
      See all 65 Publications for this product

      Publishing research using ab213? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 20 Abreviews or Q&A

      Western blot abreview for Anti-Rad51 antibody [14B4]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      20 µg
      Gel Running Conditions
      Non-reduced Denaturing (15%)
      Sample
      Human Cell lysate - whole cell (293T)
      Specification
      293T
      Blocking step
      Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 15 2015

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Rad51 antibody [14B4]

       Good
      Abreviews
      This product is known to not work in this application or species.
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
      Sample
      Human Cell (293T)
      Specification
      293T
      Permeabilization
      Yes - 0.5% Triton X100
      Fixative
      Paraformaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Jan 13 2015

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Rad51 antibody [14B4]

       Average
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Blocking step
      Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: RT°C
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: pH9
      Sample
      Human Tissue sections (Lovo Xenograft)
      Specification
      Lovo Xenograft
      Permeabilization
      No
      Fixative
      Formaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Jul 17 2013

      Question

      The customer has responded as below;

      Please could we have a credit note.

      In regards to the ab58101 antibody, I think it is generally accepted that full length Sash1 runs at 170 kDa, however another band can be observed at 130kDa in some samples (see attached paper). However, using ab58101 neither the 170 kDa or 130 kDa band can be detected, whereas the Sash1 antibody from one of your competitors detects both bands in the same sample.

      Thanks for your help and hope to hear from you soon

      Kind regards,

      Read More
      Answer

      Thank you for contacting us.

      I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

      (1) Redeemed against the original invoice if this hasn't already been paid.
      (2) Held on the account for use against a future order.
      (3) A full refund can be offered where no other invoices are outstanding.

      Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

      To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

      The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

      I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

      Read More

      Question

      Please see below for the response from the customer

      I have attached western images for the SETBP1 and Sash1 antibodies. Unfortunately I didn't save any of the IF images for the Rad51 antibody as the quality was too bad.

      Thanks!

      Kind regards,

      Read More
      Answer

      Thank you for your email.

      I would be happy to process refunds for ab213 and ab68715. Could you however let me know why the customer expecting a 170kDa band for ab58101 which should be nearly 130-140 kDa.

      Thanks!

      Read More

      Question

      Please see below for an email we have received from a customer;

      I've ordered several antibodies from AbCam via Sapphire in the last few months and on the whole the quality has been very good. However, we have had three antibodies that have not worked at all for western blot or immunofluorescence and were wondering whether we could get a refund on these three?

      Ab213 Rad51 antibody – this antibody shows a lot of background and cytoplasmic staining on immunofluorescence at the recommended concentration and lower and we are unable to see DNA repair foci following DNA damage induction via IR.

      Sapphire Order #: PO-15316
      Abcam Invoice #: 1348460E

      AB68715 SETBP1 antibody – we are unable to detect a band of the expected size of around 170 kDa on western blot.

      Sapphire Order #: PO-15671

      Abcam Invoice #: 1353502-SAP002E

      AB58101 SASH1 antibody – we were unable to detect a band at the correct size of around 170 kDa via western blot or any staining via IF.

      Sapphire Order #: PO-15316

      Abcam Invoice #: 1347756F

      All of these antibodies have been trialled by at least two experienced people in our lab and western blotting and immunofluorescence are techniques we have extensive experience with. I can provide further details of the protocols used if required.

      Thanks for your help!

      Kind regards,

      Read More
      Answer

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

      I have read the details you have kindly provided and have following further questions for better understanding of the problem;

      - Could you provide us the images? we may help you with the troubleshooting.

      Thank you very much for your cooperation. I will look forward to hearing from you soon.

      Read More

      Question

      I need your help to address the problem, the customer us Abcam( cat #: ab213) primary antibody for western blotting, and the experiment didn’t work, could you help him find out what is the problem? Please see the attached for the protocol he follows ,



      Thanks a lot , your quick response will be greatly appreciated!

      Read More
      Answer

      Thank you for contacting Abcam.

      I just had a few more protocol questions for you regarding the problems you are having with ab213 in western blot.

      1 - Why was 2 different lysis buffers used in the extraction protocol? Either of them should have been suffcient on their own.

      2 - Whenyou say that the wetern blotting did not work. Does that means there was no signal, high background, weak signal?

      3 - Could you please provide the lot number of the antibody you are using?

      I look forward to your reply and helping resolve this situation.

      Read More

      Question

      Thank you so much for your help. I will sent you update of the ab63801. And the secondary antibody we used was 1:2000 for ab213.

      Read More
      Answer

      I am happy to help. Best of luck in your experiments!

      Read More

      Question

      I have used this antibody to stain MCF7 cells successfully via ICC/IF but WB with MCF7 lysate from 50ug protein extract, with primary at 1:250 and 1:500, barely showed a band after 45' exposure

      Read More
      Answer

      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 for one vial of ab63801.

      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

      ICC: sees big red blob in nucleus (correct location) MCF7 cells, treatment with mitomicine C Ab:1/500 1h secondary: 1/200 Texas red (goat anti-mouse), works well otherwise pre-extraction with 0.25% TX fixation: 3% PFA perm step: 0.5% TX no separate blocking step, but done with primary incubation in 3% BSA suggestions: use less antibody (secondary, maybe also primary), include separate blocking step, omit pre-extraction step)

      Read More
      Answer

      Thank you for contacting us. I am sorry to hear that the antibody is giving problems. As we discussed yesterday over the phone, the following suggestions might help to get rid of the red blob you are seeing in the nuclei.: 1) use less antibody (for the secondary: e.g. 1/500 or even 1/1000, maybe also for the primary: e.g. 1/1000) 2) include a separate blocking step (e.g. 3-5% BSA or 10% goat serum) 3) omit the pre-extraction step Please let me know if these if these suggestions are of help or not. If the tips do not help, please let me know your PO or order number. If the product was purchased within the last 6 months, we can discuss a free of charge replacement, credit or refund according to our Abpromise guarantee. I wish you good luck with your experiments and look forward to hear back from you.

      Read More

      1-10 of 20 Abreviews or Q&A

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com