Anti-TopBP1 抗体 (ab2402)

Reviews (15)Q&A (2)References (32)
Western blot - Anti-TopBP1 antibody (ab2402)

    Key features and details

    • Rabbit polyclonal to TopBP1
    • Suitable for: WB
    • Reacts with: Mouse, Human, African green monkey
    • Isotype: IgG

    製品の概要

    • 製品名

      Anti-TopBP1 antibody
      TopBP1 一次抗体 製品一覧

    • 製品の詳細

      Rabbit polyclonal to TopBP1

    • 由来種

      Rabbit

    • アプリケーション

      適用あり: WBmore details

    • 種交差性

      交差種: Mouse, Human, African green monkey
      交差が予測される動物種: Chimpanzee, Gorilla, Orangutan

    • 免疫原

      Synthetic peptide (Human) - which represents a portion of human Topoisomerase (DNA) II Binding Protein I encoded within exon 28.

    • 特記事項

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

    製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab2402 in the following tested applications.

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

    アプリケーション Abreviews 特記事項
    WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 200 kDa (predicted molecular weight: 173.3 kDa).

    ターゲット情報

    • 機能

      Required for DNA replication. Plays a role in the rescue of stalled replication forks and checkpoint control. Binds double-stranded DNA breaks and nicks as well as single-stranded DNA. Recruits the SWI/SNF chromatin remodeling complex to E2F1-responsive promoters. Down-regulates E2F1 activity and inhibits E2F1-dependent apoptosis during G1/S transition and after DNA damage. Induces a large increase in the kinase activity of ATR.

    • 組織特異性

      Highly expressed in heart, brain, placenta, lung and kidney.

    • 配列類似性

      Contains 8 BRCT domains.

    • 翻訳後修飾

      Phosphorylated on serine and threonine residues in response to X-ray irradiation.
      Ubiquitinated and degraded by the proteasome. X-ray irradiation reduces ubiquitination.

    • 細胞内局在

      Nucleus. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Cytoplasm, cytoskeleton, spindle pole. Chromosome. Detected on unpaired autosomes in meiotic prophase cells. Detected on X and Y chromosomes during later stages of prophase. Colocalizes with ATR and H2AFX at unsynapsed chromosome cores during prophase (By similarity). Has a uniform nuclear distribution during G phase. Colocalizes with BRCA1 at stalled replication forks during S phase. In mitotic cells it colocalizes with BRCA1 at spindle poles and centrosomes during metaphase and anaphase. Detected in discrete foci together with PML and numerous DNA repair enzymes after DNA damage by alkylating agents, UV or gamma irradiation. Localizes to sites of DNA damage in a H2AX- independent manner.
    • Information by UniProt

    • 参照データベース

      • 別名

        • DNA topoisomerase 2 binding protein 1 antibody
        • DNA topoisomerase 2-binding protein 1 antibody
        • DNA topoisomerase II binding protein 1 antibody
        • DNA topoisomerase II binding protein antibody
        • DNA topoisomerase II-beta-binding protein 1 antibody
        • DNA topoisomerase II-binding protein 1 antibody
        • DNA topoisomerase IIbeta binding protein 1 antibody
        • Hypothetical protein KIAA0259 [Fragment] antibody
        • KIAA0259 antibody
        • TOP2BP1 antibody
        • TOPB1_HUMAN antibody
        • TOPBP 1 antibody
        • TopBP1 antibody
        • Topoisomerase (DNA) II binding protein 1 antibody
        • Topoisomerase II binding protein 1 antibody
        see all

      画像

      • Western blot - Anti-TopBP1 antibody (ab2402)
        Western blot - Anti-TopBP1 antibody (ab2402)

        Lane 1: WB Cell Lysate (5% of input IP)
        Lane 2: IP control (normal rabbit IgG)
        Lane 3: IP/WB

        Conditions:
        IP: Rabbit polyclonal to TopBP1 (1 µg)
        WB: Rabbit polyclonal to TopBP1 (1/2000 dilution)
        Positive control: HeLa Cells

        Lane 1: WB Cell Lysate (5% of input IP)
        Lane 2: IP control (normal rabbit IgG)
        Lane 3: IP/WB

        Conditions:
        IP: Rabbit polyclonal to TopBP1 (1 µg)
        WB: Rabbit polyclonal to TopBP1 (1/2000 dilution)
        Positive control: HeLa Cells

      参考文献 (32)

      ab2402 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab2402 は 32 報の論文で使用されています。

      • 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
      • Kidiyoor GR  et al. ATR is essential for preservation of cell mechanics and nuclear integrity during interstitial migration. Nat Commun 11:4828 (2020). PubMed: 32973141
      • Deshar R  et al. RNF8 mediates NONO degradation following UV-induced DNA damage to properly terminate ATR-CHK1 checkpoint signaling. Nucleic Acids Res 47:762-778 (2019). PubMed: 30445466
      • Wu M  et al. A PLK1 kinase inhibitor enhances the chemosensitivity of cisplatin by inducing pyroptosis in oesophageal squamous cell carcinoma. EBioMedicine 41:244-255 (2019). PubMed: 30876762
      • Michelena J  et al. Basal CHK1 activity safeguards its stability to maintain intrinsic S-phase checkpoint functions. J Cell Biol 218:2865-2875 (2019). PubMed: 31366665
      View all Publications for this product

      レビューと Q&A

      Show All レビュー Q&A
      レビューを送る 質問を送る

      1-10 of 17 Abreviews or Q&A

      Immunoprecipitation abreview for Anti-TopBP1 antibody - ChIP Grade

       Excellent
      Abreviews
      abreview image
      Application
      Immunoprecipitation
      Immuno-precipitation step
      Protein A/G
      Sample
      Human Cell lysate - whole cell (HeLa cells)
      Specification
      HeLa cells
      Total protein in input
      30 µg
      Read More

      Abcam user community

      Verified customer

      投稿 Jan 07 2015

      Immunocytochemistry/ Immunofluorescence abreview for Anti-TopBP1 antibody - ChIP Grade

       Excellent
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Blocking step
      BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
      Sample
      Human Cell (HeLa cells)
      Specification
      HeLa cells
      Permeabilization
      Yes - 0.2% Triton-X100
      Fixative
      Paraformaldehyde
      Read More

      Abcam user community

      Verified customer

      投稿 Nov 24 2014

      Western blot abreview for Anti-TopBP1 antibody - ChIP Grade

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      40 µg
      Gel Running Conditions
      Reduced Denaturing (4-16% gradient Tris-Gly gel)
      Sample
      Mouse Cell lysate - whole cell (MEFs)
      Specification
      MEFs
      Blocking step
      LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
      Read More

      Abcam user community

      Verified customer

      投稿 Aug 01 2014

      Western blot abreview for Anti-TopBP1 antibody - ChIP Grade

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      40 µg
      Gel Running Conditions
      Reduced Denaturing (4-16% gradient Tris-Gly gel)
      Sample
      Human Cell lysate - whole cell (HeLa cells)
      Specification
      HeLa cells
      Blocking step
      LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
      Read More

      Abcam user community

      Verified customer

      投稿 Aug 01 2014

      Western blot abreview for Anti-TopBP1 antibody - ChIP Grade

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Loading amount
      25 µg
      Gel Running Conditions
      Reduced Denaturing (3-8%)
      Sample
      Human Cell lysate - whole cell (HCT116)
      Specification
      HCT116
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
      Read More

      Abcam user community

      Verified customer

      投稿 Dec 17 2013

      Immunocytochemistry/ Immunofluorescence abreview for Anti-TopBP1 antibody - ChIP Grade

       Below Average
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Xenopus laevis Cell (Nuclei generated in oocyte extract)
      Specification
      Nuclei generated in oocyte extract
      Fixative
      Formaldehyde
      Permeabilization
      Yes - PBS-0.1% NP40
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
      Read More

      Dr. Andrew Lane

      Verified customer

      投稿 Apr 11 2013

      Western blot abreview for Anti-TopBP1 antibody - ChIP Grade

       Poor
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Xenopus laevis Cell lysate - whole cell (Xenopus egg extract)
      Loading amount
      40 µg
      Specification
      Xenopus egg extract
      Gel Running Conditions
      Reduced Denaturing (8%)
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
      Read More

      Dr. Andrew Lane

      Verified customer

      投稿 Apr 11 2013

      Immunocytochemistry/ Immunofluorescence abreview for Anti-TopBP1 antibody - ChIP Grade

       Good
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      African Green Monkey Cell (CV-1, Epithel Kidney)
      Specification
      CV-1, Epithel Kidney
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - Triton
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
      Read More

      Abcam user community

      Verified customer

      投稿 Jan 28 2013

      Question

      I complete the IP questionnaire. I want to know if you are able to identify who is the band under TopBP1 in the input and in the IP. I attach apicture of IP/wester blot. I hope that you can help me

      thank you

      Abcam IP Questionnaire
      1) Abcam product code ab2402

      2) Abcam order reference number or product batch number #1109783

      3) Description of the problem we identify an interaction between our protein of study and TopBP1 but there is another interaction more specific then TopBP1 recognized by TopBP1 antibody also in input of hela cell lines extract. The IP to TopBP1 is ok but I want to know what is the other band. The MW of Claspin is 150Kda but it migrates at 250Kda because of its iso-electric point and Chk1 is 55Kda

      4) Sample preparation:

      Type of sample whole Hela cell lysates

      Lysis buffer: Hepes-NaOH 50mM ph 7,5 , NaCl 150mM, 1% glicerol, 1% Triton X100, MgCl2 1,5mM, EGTA 5mM

      Protease inhibitors:protease inhibitor cocktail 78410 as recipe

      Phosphatase inhibitors phosphatase inhibitor cocktail 1861277 as recipe

      Amount of Lysate IPed ug or number of cells 2mg of lysate for IP

      Lysate precleared with matrix : yes

      Positive control whole Hela cell lysates 50ug

      Negative control No IP only matrix


      5) IP antibody:

      Amount/concentration or dilution used 2ug of antibody for IP

      Conjugation:

      Antibody noncovalently bound to matrix before incubation with lysate sample? NO

      Antibody noncovalently bound to matrix after incubation with lysate sample? YES

      Antibody covalently bound to matrix - please describe briefly? NO

      We used to pre-clear the cell lysates for 1h at 40C on roacking with 30ul of beads then pellet it for 5’ at 2000rpm 40C, collect the supernatant and put it to contct with antibodies O.N. Next day we used 20ul of beads/IP to bound the immunocomplex

      Antibody-lysate incubation time: Over night

      Incubation temperature 40C on roacking

      6) IP Matrix (e. beads):

      Type of matrix Protein G sepharose 4 fast flow GE Healthcare 17-0618-01

      Amount of matrix 20ul

      7) Washing after antibody-lysate incubation:

      Buffer: : Hepes-NaOH 50mM ph 7,5 , NaCl 150mM, 1% glicerol, 1% Triton X100, MgCl2 1,5mM, EGTA 5mM

      Number of washes 5

      8) IP analysis:

      Verification method: e.g reprobe in western blotting NO

      Describe verification method briefly:

      We used ECL Plus Western Blotting Detection Reagents Amersham

      OR if Western blotting reprobe performed please indicate the following:

      Reducing agent Bme

      Boiling for ≥5 min? yes

      Protein loaded ug/lane or cells/lane 50ug for input

      Percentage of gel 10%

      Type of membrane Amersham Hybond ECL Nitrocellulose Membrane

      Protein transfer verified: YES, Ponceau

      Blocking agent and concentration 5% milk in TBS+1% Tween

      Blocking time 1h

      Blocking temperature RT

      Primary antibody (If more than one was used, describe in “additional notes”) :TopBP1 ab2402

      Concentration or dilution 1:1000

      Diluent buffer 5% milk in TBS+1% Tween

      Incubation time Oven night

      Incubation temperature: 40C

      Secondary antibody:

      Species: Goat

      Reacts against: Rabbit

      Concentration or dilution 1:3000

      Diluent buffer 5% milk in TBS+1% Tween

      Incubation time 45’

      Incubation temperature: RT

      Fluorochrome or enzyme conjugate: horseradish peroxidase conjugated

      Washing after primary and secondary antibodies:

      Buffer TBS+1% Tween

      Number of washes 5

      Detection method

      Was the method successfully used to detect protein from non-IPed samples? Yes this methods detect the band of TopBP1 and another with smaller MW

      Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

      Read More
      Answer

      Thank you for your enquiry regardingab2402 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

      We regret to inform you that we do not know the identity of the band in question. We only know the identity for the target band TopBP1.

      Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not? If you have not done yet, I would advise you to check it.

      I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

      Read More

      Question

      Dear Abcam,
      I have a technical problem with a TopBP1 ab2402. I study the involment of novel protein in DNA damage response. I use a IP technic to identify if my protein of interest is a TopBP1 partner. I think that the interaction is ok but in the film there is another interaction more specific then with TopBP1. Your antibody recognized this other band with equal specificity to TopBP1. I know that your antibody can cross-react with CLASPIN (150 Kda) or CHK1 (55 Kda) but the MW is enought different. I enclose you also a photo of film with a detailed informations.
      I can hope you can help me in resolving this problem
      Best regards

      Read More
      Answer

      Thank you for contacting us.

      I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

      In order to better understand the problem, I would like to send you a questionnaire with some questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. Any images are highly appreciated.

      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. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.

      I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

      Read More

      1-10 of 17 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