Overview

  • Product name

    Anti-Securin antibody [EPR3240]
    See all Securin primary antibodies
  • Description

    Rabbit monoclonal [EPR3240] to Securin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Securin aa 100-200 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Daudi, HCT 116 lysates and HeLa cell lysates. Human testis tissue. IHC-P: human breast carcinoma tissue. ICC/IF: HeLa cells. FC: HeLa cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab79546 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 1/50.

For unpurified use at 1/100 - 1/250.

WB 1/5000 - 1/20000. Detects a band of approximately 28 kDa (predicted molecular weight: 22 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Regulatory protein, which plays a central role in chromosome stability, in the p53/TP53 pathway, and DNA repair. Probably acts by blocking the action of key proteins. During the mitosis, it blocks Separase/ESPL1 function, preventing the proteolysis of the cohesin complex and the subsequent segregation of the chromosomes. At the onset of anaphase, it is ubiquitinated, conducting to its destruction and to the liberation of ESPL1. Its function is however not limited to a blocking activity, since it is required to activate ESPL1. Negatively regulates the transcriptional activity and related apoptosis activity of TP53. The negative regulation of TP53 may explain the strong transforming capability of the protein when it is overexpressed. May also play a role in DNA repair via its interaction with Ku, possibly by connecting DNA damage-response pathways with sister chromatid separation.
  • Tissue specificity

    Expressed at low level in most tissues, except in adult testis, where it is highly expressed. Overexpressed in many patients suffering from pituitary adenomas, primary epithelial neoplasias, and esophageal cancer.
  • Sequence similarities

    Belongs to the securin family.
  • Developmental stage

    Low level during G1 and S phases. Peaks at M phase. During anaphase, it is degraded.
  • Domain

    The N-terminal destruction box (D-box) acts as a recognition signal for degradation via the ubiquitin-proteasome pathway.
    The TEK-boxes are required for 'Lys-11'-linked ubiquitination and facilitate the transfer of the first ubiquitin and ubiquitin chain nucleation. TEK-boxes may direct a catalytically competent orientation of the UBE2C/UBCH10-ubiquitin thiolester with the acceptor lysine residue.
  • Post-translational
    modifications

    Phosphorylated at Ser-165 by CDK1 during mitosis.
    Phosphorylated in vitro by ds-DNA kinase.
    Ubiquitinated through 'Lys-11' linkage of ubiquitin moieties by the anaphase promoting complex (APC) at the onset of anaphase, conducting to its degradation. 'Lys-11'-linked ubiquitination is mediated by the E2 ligase UBE2C/UBCH10.
  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • AW555095 antibody
    • C87862 antibody
    • Cut2 antibody
    • EAP 1 antibody
    • EAP1 antibody
    • ESP1 associated protein 1 antibody
    • Esp1-associated protein antibody
    • hPTTG antibody
    • MGC126883 antibody
    • MGC138276 antibody
    • Pds1 antibody
    • Pituitary tumor transforming 1 antibody
    • Pituitary tumor transforming protein 1 antibody
    • Pituitary tumor-transforming 1, isoform CRA_a antibody
    • Pituitary tumor-transforming 1, isoform CRA_b antibody
    • Pituitary tumor-transforming gene 1 antibody
    • Pituitary tumor-transforming gene 1 protein antibody
    • PTTG 1 antibody
    • PTTG antibody
    • PTTG1 antibody
    • PTTG1 protein antibody
    • PTTG1_HUMAN antibody
    • Pttg3 antibody
    • Securin antibody
    • Tumor transforming 1 antibody
    • Tumor transforming protein 1 antibody
    • Tumor-transforming protein 1 antibody
    • TUTR 1 antibody
    • TUTR1 antibody
    see all

Images

  • All lanes : Anti-Securin antibody [EPR3240] (ab79546) at 1/1000 dilution (Purified)

    Lane 1 : Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates
    Lane 2 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 22 kDa
    Observed band size: 28 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-Securin antibody [EPR3240] (ab79546) at 1/20000 dilution ((unpurified))

    Lane 1 : Daudi cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 22 kDa
    Observed band size: 28 kDa why is the actual band size different from the predicted?
    Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Securin with purified ab79546 at 1/50 dilution (4.74 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling Securin with purified ab79546 at 1/200 dilution (1.19 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Securin with purified ab79546 at 1/30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Unpurified ab79546 at 1/100 dilution staining Securin in paraffin-embedded human testis tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa cells labeling securin with unpurified ab79546 at 1/250 dilution. The cells were permeabilised with 0.1% Triton X-100. Anti-rabbit Alexa Fluor® 488 (ab150077) at 1/500 dilution was used as the secondary antibody (green). The nuclear counter stain is DAPI (blue).

  • Flow cytometric analysis of permeabilized Jurkat cells using unpurified ab79546 (red) or a rabbit IgG (negative) (green).

References

This product has been referenced in:

  • Wu MS  et al. CDC20 and its downstream genes: potential prognosis factors of osteosarcoma. Int J Clin Oncol 24:1479-1489 (2019). Read more (PubMed: 31278532) »
  • Konishi M  et al. Quantitative analyses of the metaphase-to-anaphase transition reveal differential kinetic regulation for securin and cyclin B1. Biomed Res 39:75-85 (2018). Read more (PubMed: 29669986) »
See all 12 Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Mouse Cell (Neurons)
Specification
Neurons
Permeabilization
Yes - 0.5% Triton X100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 30 2014

Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Loading amount
40 µg
Specification
Liver
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 15 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
30 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

Abcam user community

Verified customer

Submitted Jun 06 2011

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