Overview

  • Product nameAnti-Claspin antibody
    See all Claspin primary antibodies
  • Description
    Rabbit polyclonal to Claspin
  • Tested applicationsSuitable 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

  • FunctionRequired 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 similaritiesBelongs to the claspin family.
  • DomainThe 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 localizationNucleus.
  • 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

Anti-Claspin antibody images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Claspin with ab3720 at 1/1000 (1µg/ml). Detection: DAB.
  • 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

  • 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 for Anti-Claspin antibody (ab3720)

This product has been referenced in:
  • 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) »
  • 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 . Read more (PubMed: 20048340) »

See all 10 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Aug 08 2013

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...

Read More
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
Username

Abcam user community

Verified customer

Submitted Nov 07 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Aug 13 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Mr. Paul Eve

Verified customer

Submitted Aug 07 2009

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

Dr. Kirk McManus

Verified customer

Submitted May 28 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"