AnaSpec Introduces Sortase A Activity Assay Kit

AnaSpec is pleased to announce the release of a new SensoLyte® assay kit for the measurement of Sortase A activity – the SensoLyte® 520 Sortase A Activity Assay Kits.

Sortases are a family of membrane–anchored transpeptidases expressed by Gram–positive bacteria⁽¹⁾. Sortase A catalyzes the cleavage of C-terminal recognition motif (LPXTG) of multiple structurally unrelated proteins followed by formation of an amide bond with the peptidoglycan layer of the bacteria⁽²⁾. Since sortases are widely distributed among a variety of bacterial pathogens and required for virulence, they represent a promising therapeutic target for the development of novel anti-infective agents⁽³⁾. Sortase A has also been used as a biotechnology tool for a variety of protein modifications and immobilization by sortase-mediated protein ligation ⁽⁴⁾.

The SensoLyte® 520 Sortase A Assay Kit employs a novel internally quenched 5-FAM/QXL® FRET (Fluoresence Resonance Energy Transfer) substrate for the detection of protease activity. The enzyme cleaves the FRET substrate into two separate fragments resulting in the release of 5-FAM fluorescence, monitored at excitation/emission= 490/520 nm. The long wavelength fluorescence of 5-FAM is less interfered by the autofluorescence of components in biological samples and test compounds resulting in higher assay sensitivity. Assays are performed in a convenient 96-well microplate format and can detect as low as 20 ng/mL active Sortase A.

References:
1. Spirig, T. et al. Mol Microbiol 82, 1044 (2011).
2. Marraffini, LA. et al. Microbiol Mol Biol Rev 70, 192 (2006).
3. Maresso, AW. et al. Pharmacol Rev 60, 128 (2008).
4. Proft, T. Biotechnol Lett 32, 1 (2010).