Anti-TopBP1 抗体 (ab2402)

Reviews (15)Q&A (2)References (30)

Key features and details

  • Rabbit polyclonal to TopBP1
  • Suitable for: WB, IHC-P
  • 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: WB, IHC-Pmore 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.

  • ポジティブ･コントロール

    • HeLa cells and HCT116

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 1/1000 - 1/10000. Detects a band of approximately 200 kDa (predicted molecular weight: 173.3 kDa).
IHC-P Use a concentration of 4 µg/ml.

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

    参考文献

    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 30 報の論文で使用されています。

    • 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
    • Leimbacher PA  et al. MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis. Mol Cell 74:571-583.e8 (2019). PubMed: 30898438
    • Fragkos M  et al. Dicer prevents genome instability in response to replication stress. Oncotarget 10:4407-4423 (2019). PubMed: 31320994
    View all Publications for this product

    レビューと Q&A

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    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.

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    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

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