Anti-Claspin antibody (ab3720)

Reviews (5)Q&A (1)References (14)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claspin antibody (ab3720)

    Key features and details

    • Rabbit polyclonal to Claspin
    • Suitable for: IHC-P
    • Reacts with: Human
    • Isotype: IgG

    Overview

    • Product name

      Anti-Claspin antibody
      See all Claspin primary antibodies

    • Description

      Rabbit polyclonal to Claspin

    • Host species

      Rabbit

    • Tested Applications & Species

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

    • Immunogen

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

    • General notes

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

      One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

      Learn more here.

    Properties

    Applications

    The Abpromise guarantee

    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.

    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
    All applications
    Chimpanzee
    Gorilla
    Orangutan
    Application Abreviews Notes
    IHC-P
    1/500 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
    Notes
    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.

      References (14)

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

      ab3720 has been referenced in 14 publications.

      • Kobayashi G  et al. Claspin overexpression is associated with high-grade histology and poor prognosis in renal cell carcinoma. Cancer Sci 111:1020-1027 (2020). PubMed: 31912588
      • Rai R  et al. The Replisome Mediates A-NHEJ Repair of Telomeres Lacking POT1-TPP1 Independently of MRN Function. Cell Rep 29:3708-3725.e5 (2019). PubMed: 31825846
      • 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 . PubMed: 27187580
      • Ho H  et al. RhoJ regulates melanoma chemoresistance by suppressing pathways that sense DNA damage. Cancer Res 72:5516-28 (2012). PubMed: 22971344
      • Yoshida K  et al. CDC6 interaction with ATR regulates activation of a replication checkpoint in higher eukaryotic cells. J Cell Sci 123:225-35 (2010). WB ; Human . PubMed: 20048340
      View all Publications for this product

      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
      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
      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
      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
      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
      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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