Anti-Claspin antibody (ab3720)

Reviews (5)Q&A (1)References (12)

Overview

  • Product name
    Anti-Claspin antibody
    See all Claspin primary antibodies
  • Description
    Rabbit polyclonal to Claspin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Gorilla, Orangutan
  • Immunogen

    Synthetic peptide (Human) - which represented a portion of human Claspin encoded in part by exons 11, 12 and 13.

  • Positive control
    • HeLa cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab3720 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
ICC/IF Use at an assay dependent concentration. PubMed: 19124652
IP Use at 1-5 µg/mg of lysate.
WB 1/500 - 1/2500. Detects a band of approximately 175 kDa (predicted molecular weight: 150 kDa).
IHC-P 1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function
    Required for checkpoint mediated cell cycle arrest in response to inhibition of DNA replication or to DNA damage induced by both ionizing and UV irradiation. Adapter protein which binds to BRCA1 and the checkpoint kinase CHEK1 and facilitates the ATR-dependent phosphorylation of both proteins. Can also bind specifically to branched DNA structures and may associate with S-phase chromatin following formation of the pre-replication complex (pre-RC). This may indicate a role for this protein as a sensor which monitors the integrity of DNA replication forks.
  • Sequence similarities
    Belongs to the claspin family.
  • Domain
    The C-terminus of the protein contains 3 potential CHEK1-binding motifs (CKB motifs). Potential phosphorylation sites within CKB motif 1 and CKB motif 2 are required for interaction with CHEK1.
  • Post-translational
    modifications
    Phosphorylated. Undergoes ATR-dependent phosphorylation by CHEK1 during activation of DNA replication or damage checkpoints. Phosphorylation by CSNK1G1/CK1 promotes CHEK1 binding.
    Ubiquitinated by the anaphase promoting complex/cyclosome (APC/C) during G1 phase, leading to its degradation by the proteasome. Ubiquitination is mediated via its interaction with FZR1/CDH1. Following DNA damage, it is deubiquitinated by USP28 in G2 phase, preventing its degradation.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
    • Alternative names
      • Claspin antibody
      • claspin homolog (Xenopus laevis) antibody
      • Claspin homolog antibody
      • CLSPN antibody
      • CLSPN protein antibody
      • CLSPN_HUMAN antibody
      • hClaspin antibody
      • Hu Claspin antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claspin antibody (ab3720)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claspin antibody (ab3720)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Claspin with ab3720 at 1/1000 (1µg/ml). Detection: DAB.
    • Immunocytochemistry/ Immunofluorescence - Anti-Claspin antibody (ab3720)
      Immunocytochemistry/ Immunofluorescence - Anti-Claspin antibody (ab3720)This image is courtesy of an Abreview submitted by Dr Kirk McManus
      ab3720 (1/200) staining Claspin in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% TritonX100/PBS and counterstained with DAPI in order to highlight the nucleus. Please refer to abreview for further experimental details.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-Claspin antibody (ab3720)
      Immunocytochemistry/ Immunofluorescence - Anti-Claspin antibody (ab3720)
      ICC/IF image of ab2861 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2861, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References

    This product has been referenced in:
    • Liang X  et al. Anti-TGF-ß Antibody, 1D11, Ameliorates Glomerular Fibrosis in Mouse Models after the Onset of Proteinuria. PLoS One 11:e0155534 (2016). IHC . Read more (PubMed: 27187580) »
    • Ho H  et al. RhoJ regulates melanoma chemoresistance by suppressing pathways that sense DNA damage. Cancer Res 72:5516-28 (2012). Read more (PubMed: 22971344) »
    See all 12 Publications for this product

    Publishing research using ab3720? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-Claspin antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Sample
    Human Cell lysate - nuclear (HeLa nuclear cell lysate)
    Specification
    HeLa nuclear cell lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 08 2013

    Question

    I recently bought this ab, which, as written in the datasheet, detects a band of approximately 175 kDa. I completed a western blot analysis in Whole human cell lysate (MDA-MB-231 breast cancer cells) treated with increasing doses of the proteasomal inhibitor MG-132 and detected various bands; ˜270, ˜240, ˜175, ˜160, ˜125, ˜100 and ˜70 KDa. Previously, I was using an anti-Claspin antibody (from another company) that was detecting a band at ˜220kDa in these cell line. Since my strongest bands are the 270 and 240 kDa, I was wondering whether this is the actual claspin band(s) and not the 175/160 kDa?

    Also, has anybody else reported detecting two bands? It seems that I detect a lot of double bands.

    ·

    Read More
    Answer

    The predicted MW of the protein is 151 kDa for isoform 1. Two other isoforms exist, isoform 2 (144 kDa) and isoform 3 (150 kDa) (see http://www.uniprot.org/uniprot/Q9HAW4), but I am not sure under which conditions you would be able to detect either one and in which samples they might be present. The literature might be able to help in this regard.

    As for your WB results, the band should be around 175 - 180 kDa that's where the lab and other customers detected the band (see Abreviews). I cannot find an explanation for the 2 bands around 250 kDa as well as the other bands you see around 130, 100 and 70 kDa. I believe these must be non-specific bands.

    Read More

    Western blot abreview for Anti-Claspin antibody

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK 293)
    Loading amount
    20 µg
    Specification
    HEK 293
    Gel Running Conditions
    Non-reduced Denaturing (7.5)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 07 2011

    Immunoprecipitation abreview for Anti-Claspin antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HT29 tumour cell line)
    Total protein in input
    500 µg
    Specification
    HT29 tumour cell line
    Treatment
    4Gy Ionising Radiation
    Immuno-precipitation step
    Protein A/G
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 13 2009

    Western blot abreview for Anti-Claspin antibody

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (ht29 tumour cell line)
    Loading amount
    50 µg
    Specification
    ht29 tumour cell line
    Treatment
    4Gy Ionising Radiation
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Mr. Paul Eve

    Verified customer

    Submitted Aug 07 2009

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Claspin antibody

     Average
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% TritonX100 in PBS
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    Submitted May 28 2009

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