Thrombin, a serine protease, plays a central role in hemostasis
by converting soluble plasma fibrinogen into an insoluble fibrin clot and by
promoting platelet aggregation.1 Thrombin also influences a number
of normal and pathological processes, including inflammation,2 tissue
repair,3 embryogenesis,4 angiogenesis,5 and
AnaSpec is pleased to introduce the industry’s first FRET based thrombin assay
– the SensoLyte®
520 Thrombin Activity Assay Kit. This kit employs a novel internally quenched
5-FAM/QXL® 520 FRET substrate (Figure 1).
Thrombin cleaves the FRET substrate into two separate fragments resulting in
release of the fluorescence of 5-FAM, a green dye. Increase of
fluorescence can be monitored at the commonly used excitation and emission
wavelengths of 490 nm and 520 nm, respectively. The long wavelength fluorescence
of 5-FAM is less interfered by the autofluorescence of components in biological
samples and test compounds, resulting in a high signal to background ratio.
Figure 2 shows the inhibition data.
||Figure 1. Proteolytic cleavage of the 5-FAM/QXL®
520 FRET peptide by thrombin increases the distance between the donor and
quencher. This results in the release of 5-FAM fluorescence.
||Figure 2. Inhibition of thrombin activity was determined
using the SensoLyte® 520 Thrombin Assay Kit.
AnaSpec also offers a shorter wavelength thrombin assay, which employs
an AFC (7-amido-4-trifluoromethylcoumarin) labeled
substrate, the SensoLyte®
AFC Thrombin Activity Assay Kit.
Related Products :
and Related Peptides
JT. et al. J. Thromb. Hemost. 5, 95(2007).
K. et al. Curr. Opin. Hematol. 14, 55 (2007).
M. et al. Exp. Dermatol. 15, 523 (2006).
CT. et al. Science 293, 1666 (2001).
ME. et al. Biochem. Soc. Trans. 30, 173 (2002).
KM. et al. Semin. Thromb. Hemost. 34, 734 (2008).