Overview

  • Product name

    Mouse Thrombin ELISA Kit
    See all Thrombin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 6.3%
  • Sample type

    Serum, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    17.5 pg/ml
  • Range

    0.31 ng/ml - 20 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 98 94% - 104%
    Cell culture media 99 92% - 103%
    Hep Plasma 110 106% - 112%
    EDTA Plasma 108 95% - 117%
    Cit plasma 97 91% - 102%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Thrombin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Thrombin protein in mouse serum, plasma, cell culture supernatants, and tissue extracts.


    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.


    Thrombin (activated Factor II [IIa]) is a coagulation protein that has many effects in the coagulation cascade. Thrombin is a serine protease that converts soluble fibrinogen into insoluble strands of fibrin, as well as catalyzing many other coagulation-related reactions. Thrombin is in the form of alpha-thrombin that is the immediate end-product of prothrombin activation: two further thrombin products can be identified, beta- and gamma-thrombin. These are degraded forms that may arise from autodigestion of a thrombin preparation.


    Sensitivity:


    Serum, plasma and cell culture supernatant - 36 pg/mL


    Tissue extracts - 17.5 pg/mL

  • 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 Mouse Thrombin Capture Antibody 1 x 600µl
    10X Mouse Thrombin 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 4BR 1 x 6ml
    Mouse Thrombin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Thrombin, which cleaves bonds after Arg and Lys, converts fibrinogen to fibrin and activates factors V, VII, VIII, XIII, and, in complex with thrombomodulin, protein C. Functions in blood homeostasis, inflammation and wound healing.
  • Tissue specificity

    Expressed by the liver and secreted in plasma.
  • Involvement in disease

    Factor II deficiency
    Ischemic stroke
    Thrombophilia due to thrombin defect
    Pregnancy loss, recurrent, 2
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
    Contains 2 kringle domains.
    Contains 1 peptidase S1 domain.
  • Post-translational
    modifications

    The gamma-carboxyglutamyl residues, which bind calcium ions, result from the carboxylation of glutamyl residues by a microsomal enzyme, the vitamin K-dependent carboxylase. The modified residues are necessary for the calcium-dependent interaction with a negatively charged phospholipid surface, which is essential for the conversion of prothrombin to thrombin.
    N-glycosylated. N-glycan heterogeneity at Asn-121: Hex3HexNAc3 (minor), Hex4HexNAc3 (minor) and Hex5HexNAc4 (major). At Asn-143: Hex4HexNAc3 (minor) and Hex5HexNAc4 (major).
  • Cellular localization

    Secreted, extracellular space.
  • Information by UniProt
  • Alternative names

    • Coagulation factor II
    • Coagulation factor II thrombin
    • EC 3.4.21.5
    • F2
    • Factor II
    • Prepro coagulation factor II
    • Prothrombin
    • Prothrombin B chain
    • PT
    • RPRGL2
    • Serine protease
    • THPH1
    • THRB_HUMAN
    • Thrombin heavy chain
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab230933 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.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Thrombin were measured in duplicate, interpolated from the Thrombin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:12000, plasma (citrate) 1:2000, plasma (EDTA) 1:500, and plasma (heparin) 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombin concentration was determined to be 37.1 µg/mL in serum, 3.7 µg/mL in plasma (citrate), 5.4 ug/mL in plasma (EDTA), and 4.48 µg/mL in plasma (heparin).

  • The concentrations of Thrombin were measured in duplicate and interpolated from the Thrombin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombin concentration was determined to be 2.29 ng/mL in mouse liver extract.

Protocols

References

This product has been referenced in:

  • Kanki H  et al. ß-arrestin-2 in PAR-1-biased signaling has a crucial role in endothelial function via PDGF-ß in stroke. Cell Death Dis 10:100 (2019). Read more (PubMed: 30718498) »
See 1 Publication for this product

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