Overview

  • Product name

    Human Securin ELISA Kit
    See all Securin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Securin 5 6.4%
    Inter-assay
    Sample n Mean SD CV%
    Securin 3 5.2%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    64 pg/ml
  • Range

    102 pg/ml - 6500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 98 89% - 105%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Securin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Securin protein in Human  cell extract


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Securin (PTTG1) is a regulatory protein during the metaphase-anaphase transition of the cell cycle. Before the transition, Securin is bound to Separase in the cytoplasm. During the initiation of anaphase, Securin is dephosphorylated and is then recognized by the APC-Cdc20 complex. This leads to the ubiquination and degradation of Securin and the release of Separase. Free Separase is then able to start the process of chromosomal separation.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Securin Capture Antibody 1 x 600µl
    10X Human Securin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPR 1 x 6ml
    Human Securin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
  • Alternative names

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

Associated products

Applications

Our Abpromise guarantee covers the use of ab230935 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The Securin standard curve was prepared as described in Section 10.

  • The concentrations of Securin were measured in duplicate and interpolated from the Securin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Securin concentration was determined to be 4123 pg/mL in Daudi Cell Extract, 2086 pg/ml in U937 Cell Extract, and 2017 pg/mL in HUVEC Cell Extract.

Protocols

References

ab230935 has not yet been referenced specifically in any publications.

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