AnaSpec, EGT Group is pleased to announce the release of the industry first fluorimetric Factor Xa activity assays - The SensoLyte^® 520 Factor Xa Assay Kit and the SensoLyte^® Rh110 Factor Xa Assay Kit. The SensoLyte^® 520 Factor Xa Assay Kit employs a FRET (5-FAM/QXL™ 520) substrate and the SensoLyte^® Rh110 Factor Xa Assay Kit, a substrate labeled with the fluorophore, Rh110. Both substrates are fluorescent substrates emitting in the green range of 520 nm, with excitation at 490 nm. The longer-wavelength spectra of 5-FAM and Rhodamine 110 provide greater sensitivity and less interference from other reaction components, making these kits ultra-sensitive. The detection limit of the SensoLyte^® 520 kit can reach as low as 7.8 ng/mL while the SensoLyte^® Rho110 reaches as low as 0.31 ng/mL.

• Kits optimized to detect Factor Xa enzyme activity

• Fluorometric, long wavelength (Ex/Em=490/520 nm) read-out assays

• Minimal autofluorescence

• Kits provide ample reagents to perform 100 assays in a 96-well plate format

• Assays can be completed in one hour

Factor Xa (FXa) is a serine endopeptidase composed of two disulfide-linked subunits. FXa leads to blood clot formation by converting prothrombin to thrombin through the prothrombinase complex. FXa is generated from zymogen Factor X via the intrinsic and extrinsic pathways and is the rate-limiting step in the propagation of thrombin generation. In the presence of Ca²⁺ ions, FXa forms prothrombinase with factor Va on the phospholipid membrane of the activated platelets.^1-3 FXa has emerged as an attractive target for drug discovery for thromboembolic diseases.^4,5

Figure 1. Inhibition of Factor Xa activity as measured with SensoLyte^® Rh110 Factor Xa Assay Kit.

Figure 2. Inhibition of Factor Xa activity as measured with the SensoLyte^® 520 Factor Xa Assay Kit.

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Reference

1. Furie, B. and Furie, BC. Cell 53:505–518 (1988).
2. Davie, EW. et al. Biochemistry 30:10363–10370(1991).
3. Mann, KG. Thromb Haemost 82:165-174 (1999).
4. Camm, AJ. and Bounameaux, H. Drugs 71:1503-1526 (2011).
5. Weitz, JI. Thromb Res 127 Suppl 2:S5-S12 (2011).