Anti-Rad9 antibody (ab70810)

Reviews (1)Q&A (2)References (6)
Western blot - Anti-Rad9 antibody (ab70810)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)
  • Immunoprecipitation - Anti-Rad9 antibody (ab70810)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)

Key features and details

  • Rabbit polyclonal to Rad9
  • Suitable for: IHC-P, WB, IP
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Rad9 antibody

  • Description

    Rabbit polyclonal to Rad9

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to a region between residue 350 and the C-terminus (residue 391) of Human Rad9 (NP_004575.1)

  • Positive control

    • Whole cell lysate from HeLa and 293T cells.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab70810 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
IP
Human
WB
Human
All applications
Cow
Dog
Pig
Ferret
Rhesus monkey
Gorilla
Bat
Application Abreviews Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
1/2000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 43 kDa).
IP
Use at 2-5 µg/mg of lysate.
Notes
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB
1/2000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 43 kDa).
IP
Use at 2-5 µg/mg of lysate.

Target

  • Function

    Component of the 9-1-1 cell-cycle checkpoint response complex that plays a major role in DNA repair. The 9-1-1 complex is recruited to DNA lesion upon damage by the RAD17-replication factor C (RFC) clamp loader complex. Acts then as a sliding clamp platform on DNA for several proteins involved in long-patch base excision repair (LP-BER). The 9-1-1 complex stimulates DNA polymerase beta (POLB) activity by increasing its affinity for the 3'-OH end of the primer-template and stabilizes POLB to those sites where LP-BER proceeds; endonuclease FEN1 cleavage activity on substrates with double, nick, or gap flaps of distinct sequences and lengths; and DNA ligase I (LIG1) on long-patch base excision repair substrates. RAD9A possesses 3'->5' double stranded DNA exonuclease activity. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex.

  • Sequence similarities

    Belongs to the rad9 family.

  • Post-translational
    modifications

    Constitutively phosphorylated on serine and threonine amino acids in absence of DNA damage. Hyperphosphorylated by PRKCD and ABL1 upon DNA damage. Its phosphorylation by PRKCD may be required for the formation of the 9-1-1 complex.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Cell cycle checkpoint control protein antibody
      • Cell cycle checkpoint control protein RAD9A antibody
      • DNA repair exonuclease rad9 homolog A antibody
      • hRAD 9 antibody
      • hRAD9 antibody
      • Rad 9 antibody
      • RAD 9A antibody
      • RAD9 (S pombe) homolog antibody
      • RAD9 homolog A antibody
      • RAD9 homolog antibody
      • Rad9a antibody
      • RAD9A_HUMAN antibody
      see all

    Images

    • Western blot - Anti-Rad9 antibody (ab70810)
      Western blot - Anti-Rad9 antibody (ab70810)
      All lanes : Anti-Rad9 antibody (ab70810) at 0.1 µg/ml

      Lane 1 : HeLa whole cell lysate at 50 µg
      Lane 2 : HeLa whole cell lysate at 15 µg
      Lane 3 : HeLa whole cell lysate at 5 µg
      Lane 4 : 293T whole cell lysate at 50 µg

      Developed using the ECL technique.

      Predicted band size: 43 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Rad9 with ab70810 at 1/1000 (1µg/ml). Detection: DAB.
    • Immunoprecipitation - Anti-Rad9 antibody (ab70810)
      Immunoprecipitation - Anti-Rad9 antibody (ab70810)
      1mg whole cell lysate from HeLa cells was immunoprecipitated using ab70810 at 3ug/mg of lysate (lane 1) or a control rabbit Ig (lane 2). For the subsequent western blot, 20% of the immunoprecipitate was loaded per lane, and probed with ab70810 at 1ug/ml.
      Detection: chemiluminescence with exposure time of 30 seconds.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rad9 antibody (ab70810)
      IHC image of ab70810 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab70810, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    References (6)

    Publishing research using ab70810? Please let us know so that we can cite the reference in this datasheet.

    ab70810 has been referenced in 6 publications.

    • Mooser C  et al. Treacle controls the nucleolar response to rDNA breaks via TOPBP1 recruitment and ATR activation. Nat Commun 11:123 (2020). PubMed: 31913317
    • Prati B  et al. Three Prime Repair Exonuclease 1 (TREX1) expression correlates with cervical cancer cells growth in vitro and disease progression in vivo. Sci Rep 9:351 (2019). PubMed: 30674977
    • Leimbacher PA  et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell 74:571-583.e8 (2019). PubMed: 30898438
    • Broustas CG  et al. RAD9A promotes metastatic phenotypes through transcriptional regulation of anterior gradient 2 (AGR2). Carcinogenesis 40:164-172 (2019). PubMed: 30295739
    • Gong Y  et al. PHF11 promotes DSB resection, ATR signaling, and HR. Genes Dev 31:46-58 (2017). WB ; Mouse . PubMed: 28115467
    • Panigrahi SK  et al. Regulation of NEIL1 protein abundance by RAD9 is important for efficient base excision repair. Nucleic Acids Res 43:4531-46 (2015). PubMed: 25873625

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    ChIP on chip abreview for Anti-Rad9 antibody

     Good
    Abreviews
    abreview image
    Application
    ChIP on chip
    Sample
    Human Cell lysate - nuclear (prostate cancer cells)
    Specification
    prostate cancer cells
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% formaldehyde
    Detection step
    Microarray
    Positive control
    RAD9 has been comfirmed to bind p21 promoter region, so after I precipatated DNA with RAD9 abs, I used precipiated DNA as template, with specific primers for RT-PCR and check p21 promoter region. AB70810 gave positive results as previous reported.

    Yes, Following qPCR, I did CHIP microarray with annealing CHIP'd DNA. I got excited data, and now I am doing the following research.

    other company RAD9 antibodies, INPUT
    Negative control
    mouse IgG
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    Abcam user community

    Verified customer

    Submitted Jul 31 2012

    Question

    Thanks for your great offer. Your information is very helpful to me. Yes, I would like to proceed and test ab70810 in ChIP, since my aim is to select any RAD9 antibody whic works for chip-on-chip, is it OK for you to send me another mouse monoclonal ab117848 to me, so I can test them in parallel? I will wait for your answer and follow your suggestion for the following process. Best Regards.

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    Answer

    AB70810 DISCOUNT CODE: XXXX Expiration date: AB117848 DISCOUNT CODE: XXXX Expiration date: I am very pleased to hear you would like to accept our offer and test both ab70810 and ab117848 and in ChIP. Above are the code that I have created for each testing discount. Please note the the discount code is attached to the specific antibody that you will be reviewing. These codes will give you 1 free primary antibody each before the expiration date. To redeem this offer, please submit an Abreview in ChIP and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. Thank you for your reply. I can send you 100% Abreview codes for both products. Each would require that you test and submit and Abreview before you could recieve the free primary antibody.

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    Question

    Hi, Good morning. I need do chip-on-chip experiment on a gene named RAD9A. I searched your website, Abcam has 9 RAD9A pimary antibdies, but none of them is a chip graded RAD9A primary antibody in Abcam list. My silly question: is there any possible that Abcam can make a chip graded RAD9A primary antibody? Best

    Read More
    Answer

    Thank you for contacting us. While we do not have a ChIP graded RAD9A antibody. It is highly likely that the products we do have may work in your experiment. Ideally, specific antibodies for ChIP should be affinity-purified. Polyclonal antibodies are also recommended over monoclonal antibodies. Ab70810 Anti-Rad9 antibody is an affinity purified rabbit polyclonal that has been shown to work in IP, Western blot and IHC. If you are interested in testing this antibody in ChIP on human tissue I would be able to offer you a 100% Abreview discount. Under program I can offer a discount off a future purchase if you buy ab70810 test it in ChIP submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. Details of this offer are below. Should the rabbit polyclonal not be appropriate for your experiments, ab117848 Anti-Rad9 is an affinity purified mouse monoclonal that may also be an excellent candidate for your experiments. In that case, you the 100% Abreview discount for this product. The only caveat being that polyclonal antibodies could recognize more epitopes than a monoclonal and therefore have a greater chance of working. These are the links to the two products that I have recommended: ab70810: https://www.abcam.com/Rad9-antibody-ab70810.html ab117848: https://www.abcam.com/Rad9-antibody-7B11-ab117848.html If you have not previously validated an antibody for ChIP, I would suggest that you review section 2 of the protocol booklet I've attached: "How to validate an antibody for ChIP" If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab70810 in ChIP. I will then send a discount code. This code must be issued before purchasing ab70810 so please wait for my reply before ordering. 2. Purchase ab70810 either by phone, fax, or online (www.abcam.com). 3. Test it in ChIP. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab70810 turns out to be unsuitable for ChIP you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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