Overview

  • Product nameAnti-TopBP1 antibody - ChIP Grade
    See all TopBP1 primary antibodies
  • Description
    Rabbit polyclonal to TopBP1 - ChIP Grade
  • Tested applicationsSuitable for: IP, WB, IHC-P, ChIP, ICC/IF, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Human, African Green Monkey
    Predicted to work with: Chimpanzee, Gorilla, Orangutan
  • Immunogen

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

  • Positive control
    • HeLa cells and HCT116

Properties

Applications

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.

Application Abreviews Notes
IP Use a concentration of 0.5 - 2.5 µg/ml. May cross-react with other proteins such as CHK1 or possibly claspin.
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.
ChIP Use at an assay dependent concentration. PubMed: 17913746
ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

Target

  • FunctionRequired 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.
  • Tissue specificityHighly expressed in heart, brain, placenta, lung and kidney.
  • Sequence similaritiesContains 8 BRCT domains.
  • Post-translational
    modifications
    Phosphorylated on serine and threonine residues in response to X-ray irradiation.
    Ubiquitinated and degraded by the proteasome. X-ray irradiation reduces ubiquitination.
  • Cellular localizationNucleus. 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
  • Database links
  • Alternative names
    • 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

Anti-TopBP1 antibody - ChIP Grade images



  • Predicted band size : 173.3 kDa


    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

  • ab2402 (4µg/ml) staining TOPBP1 in human duodenum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining in the epithelium.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunofluorescence analysis of African Green Monkey CV-1 (kidney epithelial) cells, staining TopBP1 with ab2402.

    Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 3% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/500 in 1% BSA in PBS) for 16 hours at 4°C. An AlexaFluor®488-conjugated donkey anti-rabbit polyclonal IgG (1/1000) was used as the secondary antibody.

    See Abreview

References for Anti-TopBP1 antibody - ChIP Grade (ab2402)

This product has been referenced in:
  • Li L  et al. Targeting poly(ADP-ribose) polymerase and the c-Myb-regulated DNA damage response pathway in castration-resistant prostate cancer. Sci Signal 7:ra47 (2014). Read more (PubMed: 24847116) »
  • Choi SH  et al. TopBP1 and Claspin contribute to the radioresistance of lung cancer brain metastases. Mol Cancer 13:211 (2014). WB ; Human . Read more (PubMed: 25216549) »

See all 15 Publications for this product

Product Wall

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
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Submitted Jan 07 2015

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
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Submitted Nov 24 2014

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
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Submitted Aug 01 2014

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
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Submitted Aug 01 2014

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
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Submitted Dec 17 2013

Abcam has not validated the combination of species/application used in this Abreview.
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
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Dr. Andrew Lane

Verified customer

Submitted Apr 11 2013

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Dr. Andrew Lane

Verified customer

Submitted Apr 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Submitted Jan 28 2013

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.

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