Overview

  • Product name

    Thrombin Activity Assay Kit (Fluorometric)
    See all Thrombin kits
  • Detection method

    Fluorescent
  • Sample type

    Plasma, Purified protein
  • Assay type

    Quantitative
  • Sensitivity

    1 ng
  • Species reactivity

    Reacts with: Mammals, Other species
  • Product overview

    Thrombin Activity Assay Kit (Fluorometric) (ab197006) is simple, rapid and can detect Thrombin activity as low as 1 ng in samples.


    In the thrombin activity assay protocol, a synthetic AMC-based peptide substrate is proteolytically cleaved by thrombin to release a fluorophore, AMC, which can be easily quantified by fluorescence reader at Ex/Em = 350/450 nm.


    Thrombin activity assay protocol summary:
    - add standards and samples to wells
    - add reaction mix
    - analyze with a microplate reader for 30-60 min

  • Notes

    Thrombin enzyme (Factor IIa, EC 3.4.21.5), a serine protease, is an important clotting factor in the coagulation cascade that involves the conversion of soluble fibrinogen to insoluble active fibrin strands.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Thrombin Assay Buffer 1 x 15ml
    Thrombin Dilution Buffer 1 x 1ml
    Thrombin Enzyme Standard 1 x 5µl
    Thrombin Substrate 1 x 0.5ml
  • 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
    • F2
    • Prepro coagulation factor II
    • PT
    • RPRGL2
    • THPH1
    • THRB_HUMAN
    • Thrombin heavy chain
    see all

Images

  • Thrombin activity was measured in plasma samples in the presence and absence of aThrombin inhibitor, PPACK Dihydrochloride.  S = Substrate, I = Inhibitor, AB = Activation Buffer containing Factor Xa. Assays were performed following the kit protocol.

Protocols

References

This product has been referenced in:

  • Shefa AA  et al. Enhancement of hemostatic property of plant derived oxidized nanocellulose-silk fibroin based scaffolds by thrombin loading. Carbohydr Polym 208:168-179 (2019). Read more (PubMed: 30658788) »
  • He P  et al. Timely and large dose of clotting factor IX provides better joint wound healing after hemarthrosis in hemophilia B mice. Int J Hematol 110:59-68 (2019). Read more (PubMed: 31006077) »
See all 2 Publications for this product

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