ADAM10 has been shown to be an effective α-secretase in vitro and in vivo. This enzyme acts as a sheddase to cleave cell surface proteins involved in neuropathology, inflammatory response and tumor progression. The SensoLyte® 520 ADAM10 Activity Assay Kit is an optimized assay that can be used to detect the ADAM 10 activity. The unique FRET substrate contained in the kit was designed to reduce the cross reactivity with ADAM17 (also called tumor necrosis factor-α-converting enzyme, TACE). When active ADAM10 cleaves the FRET substrate, it results in an increase of 5-FAM fluorescence monitored at excitation/emission = 490/520 nm. The long wavelength fluorescence of 5-FAM is also less interfered by the autofluorescence of components in biological samples and test compounds. This assay can detect as low as 0.25 ng/mL active ADAM10.
Component A: 5-FAM /QXL™ 520 ADAM10 substrate Ex/Em=490 nm/520 nm upon cleavage: 1 mM, 50µL
Component B: 5-FAM fluorescence reference standard, Ex/Em=490 nm/520 nm: 1 mM, 12 µL
Component C: Human recombinant ADAM10: 0.1 mg/mL, 10 µL
Component D: Assay Buffer: 25 mL
Component E: Inhibitor: 1 mM, 10µL
Storage & stability
Store component C at -80°C. Store all other components at -20°C. Component D can be stored at room temperature for convenience. Protect components A and B from light and moisture.